Objective To investigate the current distribution of radiotherapy resources in Gansu Province, evaluate the equity of resource allocation, and provide a scientific basis for optimizing regional resource allocation. Methods A questionnaire survey was carried out to assess radiotherapy resources in medical institutions across Gansu Province, China. The equity of radiotherapy resource distribution and associated disparities were assessed using the Gini coefficient, Lorenz curve, and Theil index. Results A total of 23 medical institutions in Gansu Province provided radiotherapy services, comprising 39 radiotherapy devices and 438 professionals, of whom medical physicists accounted for 16.9%. The radiotherapy frequency was 0.47 cases per thousand population. The Gini coefficients for radiotherapy resource distribution ranged from 0.38 to 0.56 by population and from 0.52 to 0.70 by geography. The Theil index for radiotherapy resources ranged from 1.36 to 3.67. Conclusion Radiotherapy resources in Gansu Province were insufficient, and the capacity of radiotherapy service was suboptimal. The equity of radiotherapy resource allocation by geography was worse than that by population. Therefore, it is imperative to address the shortage of radiotherapy resources, strengthen the professional workforce, enhance the capacity radiotherapy service and resource utilization, optimize resource allocation, and promote regional equity in radiotherapy provision in Gansu Province.

This study aims to establish a high-performance size-exclusion chromatography (HPSEC) method for determining the content of the classical swine fever virus (CSFV) E2 protein and screen the optimal stabilizer to enhance the stability of this protein. The optimal detection conditions were determined by optimizing the composition of the mobile phase, and characteristic chromatographic peaks were identified by SDS-PAGE and Western blotting. The specificity, repeatability, precision, linearity, limit of detection (LOD), and limit of quantitation (LOQ) of the method were assessed. The method established was used to determine the content of CSFV E2 protein antigen and vaccine. Differential scanning fluorimetry (DSF) was employed to screen the buffer system, pH, and salt ion concentrations, and sugar, amino acid, and alcohol stabilizers were further screened. The results showed that using a 200 mmol/L phosphate buffer provided the best column efficiency. An antigen-specific chromatographic peak appeared at the retention time of 18 min, which was identified as the CSFV E2 protein by SDS-PAGE and Western blotting. The method exhibited high specificity for detecting the CSFV E2 protein, with no absorbance peak observed in the blank control. The relative standard deviation (RSD) of the peak area for six repeated injections of the CSFV E2 protein was 0.74%, indicating good repeatability of the method. The RSD for repeated detection of two different concentrations of CSFV E2 protein samples by different operators at different time points was less than 2%, suggesting good intermediate precision of the method. The peak area of the CSFV E2 protein was linearly related to its concentration, with the regression equation showing R² of 1.000. The LOD and LOQ of the method were 14.88 μg/mL and 29.75 μg/mL, respectively. Application of the developed method in the detection of three batches of CSFV E2 protein antigen and three batches of vaccine demonstrated results consistent with those from the bicinchoninic acid (BCA) assay, which meant that the method could accurately determine the content of CSFV E2 protein antigen and vaccine. The DSF method identified 50 mmol/L Tris-HCl at pH 8.0 as the optimal buffer, and the addition of sugar and alcohol stabilizers further improved the stability of the CSFV E2 protein. The HPSEC method established in this study is simple, fast, and exhibits good accuracy and repeatability, enabling precise measurement of the CSFV E2 protein content. It is expected to play a crucial role in the quality control of the CSFV E2 vaccine. Furthermore, the strategy for improving the CSFV E2 protein stability, identified through DSF screening, has significant implications for enhancing the stability of the CSFV E2 vaccine.
Classical Swine Fever Virus/chemistry* ; Chromatography, Gel/methods* ; Animals ; Swine ; Viral Envelope Proteins/immunology*

Objective Immune infiltration,as well as their implications across various cancers,and to establish a clinical prognostic model based on the findings.Methods The expression profile and clinical information of endometrial cancer tissue and adjacent tissues in TCGA endometrial cancer database were used,the differential expression genes were screened out for analysis,and the mRNA co-expressed by LncRNA was analyzed by GO enrichment and KEGG signaling pathway.The differential genes between endometrial cancer and adjacent tissues were intersected with the basement membrane protein genes.The selected differentially expressed genes were combined with survival status and survival time to screen out Hub genes by Lasso-cox regression analysis.Multivariate Cox regression analysis was used to establish a prognostic model,and pan-cancer analysis and immunoinfiltration correlation analysis were further performed.Results Six basal membrane protein Hub genes,ADAMTS5,EVA1C,THBS4,CTSD,ITGAV and LAMA1,were identified as associated with the prognosis of patients with endometrial cancer,and found that the survival rate of patients decreased significantly with the increase of risk score.Pan-cancer analysis found that these 6 genes were significantly different in most cancer types,and high expression in GBMLGG(glioma),LGG(low-grade glioma of the brain),LAML(acute myeloid leukemia),UVM(Uveal melanoma),ACC(adrenal cortical cancer)and other cancers had poor prognosis.The potential role of these genes in tumor immunotherapy was also explored,and significant negative correlation was found with Th 17 cells,Th2 cells,NK CD56 bright cells and other immune cells(P＜0.01),and significant positive correlation was found with Tcm,iDC,Eosinophils,aDC and other immune cells(P＜0.01).Conclusion Basal membrane protein gene has high clinical value in the diagnosis and prognosis of endometrial cancer,and can be used as a prognostic marker and potential therapeutic target for patients with endometrial cancer.

Gonorrhea, caused by Neisseria gonorrhoeae, is one of the most frequently reported infectious diseases. With the increasing antibiotic resistance in Neisseria gonorrhoeae, gonorrhea has become a major public health problem worldwide, making it imperative to develop a safe and effective vaccine. Lipooligosaccharides (LOS), which exist on the outer surface of gram-negative bacteria, contain many important antigenic determinants. In recent years, a large number of studies have shown that LOS may become the most potential target of Neisseria gonorrhoeae vaccine and immunotherapy. This article reviewed the structure of LOS, its role in Neisseria gonorrhoeae infection, research progress in LOS vaccine and the challenges faced in vaccine development, aiming to provide reference for further study.

Objective:To analyze the clinical characteristics and prognosis of patients infected with novel coronavirus Omicron variant in Shanghai, as to provide a reference for epidemic prevention, clinical diagnosis, and treatment.Methods:Altogether 4 264 novel coronavirus Omicron variant-infected patients with positive results of nucleic acid admitted to Shanghai New International Expo Center N3 Mobile Cabin Hospital from April 2 to May 7, 2022, were included. The demographic and baseline clinical characteristics, treatment strategy, prognosis, and different factors affecting the length of hospital stay were analyzed.Results:A total of 4 264 novel coronavirus variant Omicron-infected cases were collected, including 3 111 cases (73.0%) asymptomatic infections and 1 153 cases (27.0%) mild infections. The overall median age was 45 (33, 55) years old with a range from 2 years old to 81 years old. The male to female ratio was 1.37∶1. Altogether 3 305 cases (77.5%) had been vaccinated, of which 3 166 cases completed more than 2 doses. The upper respiratory tract symptoms such as cough and expectoration were the most common clinical manifestations of these infected patients. During the course of the disease, patients with asymptomatic infection were mainly treated with traditional Chinese medicine (TCM, 55.1%) and clinical observation (36.8%), and those with mild infection were mainly treated with TCM (42.2%) or integrated Chinese and Western medicine (30.4%). All patients were cured and discharged. The overall median length of hospital stay and the negative conversion time of nucleic acid were 9 (6, 10) days and 8 (5, 9) days, respectively. Compared with the asymptomatic infected patients, the hospitalization duration and the nucleic acid negative conversion time of the mildly infected patients were slightly longer [days: 10 (8, 11) vs. 9 (5, 10); 8 (6, 10) vs. 7 (4, 9), both P < 0.001]. Multiple linear regression analysis showed that the increasing age and mild infection were associated with longer hospitalization duration, and the treatment of TCM or integrated Chinese and Western medicine was associated with shortened length of hospital stay (all P < 0.05). Conclusions:The current novel coronavirus Omicron variant epidemic in Shanghai mainly caused asymptomatic and mild infections. The young and middle-aged population had a relatively high infection rate. The upper respiratory tract symptoms such as cough and expectoration were the most common clinical symptoms. Elderly and confirmed patients had prolonged hospitalization duration, while for patients receiving TCM treatment, the hospitalization duration was shortened.

Objective:To explore the effect and molecular mechanism of capsaicin receptor(TRPV1) on neuronal autophagy and depression-like behavior in mice.Methods:Using the method of random number table, 87 C57 male mice were divided into Sham operation group (Sham group), cerebral ischemia/reperfusion group (I/R group) and capsazepine(CPZ) preconditioning cerebral ischemia/reperfusion group (I/R+ CPZ group), with 28 mice in each group due to 3 incompatible.Mice in the I/R group were subjected to right middle cerebral artery occlusion (MCAO) to establish a cerebral ischemia-reperfusion model.Mice in the I/R+ CPZ group were injected with CPZ in the lateral ventricle prior to moulding.Mice in the Sham group had only wire plugs inserted and no arterial embolization was performed.The mNSS score was used to evaluate the degree of neurological deficits.The depression-like behaviour of mice was detected by the tail suspension test and forced swimming test.The infarct volume was observed by TTC staining.The pathological changes in the amygdala were observed by HE staining, and the expression of Beclin-1, LC3, p62 and p-PI3K, p-AKT and p-mTOR proteins were detected by Western blot.Statistical analysis was performed using SPSS 23.0 software.The t-test was used for comparison between two groups and one-way ANOVA was used for comparison of multiple group. Results:The neurological deficit score in I/R+ CPZ group (9.77±2.32) was significantly lower than that in I/R group (12.85±2.73) ( t=3.10, P<0.01). Compared with I/R group, the tail suspension immobility time of I/R+ CPZ Group ((93.28±50.69)s, (143.80±35.61) s; t=2.94, P<0.01) and the forced swimming immobility time ((139.50±13.33)s, (175.30±19.78)s; t=2.94, P<0.01) were significantly reduced.The results of TTC staining showed that the cerebral infarct volume in I/R+ CPZ group was significantly lower than that in I/R group ((19.30±5.19)%, (33.60±3.90)%; t=5.40, P<0.01). HE staining showed that the number of cells in the amygdala region of mice in the I/R+ CPZ group increased compared with that in the I/R group, with tighter arrangement and reduced deep staining of nuclear fixation.Western blot showed that compared with I/R group, the expression levels of autophagy related proteins Beclin-1( t=2.94, P<0.05) and LC3 ( t=3.16, P<0.05) in amygdala of I/R+ CPZ group were down-regulated, while the expression levels of p62( t=3.60, P<0.05), p-PI3K ( t=7.79, P<0.01), p-AKT ( t=4.15, P<0.01) and p-mTOR ( t=6.15, P<0.01) were up-regulated. Conclusion:Cerebral ischemia/reperfusion activates neuronal autophagy, and CPZ may regulate the PI3K-AKT-mTOR pathway, thus inhibits excessive activation of autophagy, thereby acting as a neuroprotective agent and improving post-stroke depression-like behaviour.

To rapidly and accurately manipulate genome such as gene deletion, insertion and site mutation, the whole genome of a very virulent strain Md5 of Marek's disease virus (MDV) was inserted into bacterial artificial chromosome (BAC) through homogeneous recombination. The recombinant DNA was electroporated into DH10B competent cells and identified by PCR and restriction fragment length polymorphism analysis. An infectious clone of Md5BAC was obtained following transfection into chicken embryo fibroblast (CEF) cells. Furthermore, a lorf10 deletion mutant was constructed by two step Red-mediated homologous recombination. To confirm the specific role of gene deletion, the lorf10 was reinserted into the original site of MDV genome to make a revertant strain. All the constructs were rescued by transfection into CEF cells, respectively. The successful packaging of recombinant viruses was confirmed by indirect immunofluorescence assay. The results of growth kinetics assay and plaques area measurement showed that the lorf10 is dispensable for MDV propagation in vitro. Overall, this study successfully constructed an infectious BAC clone of MDV and demonstrated its application in genome manipulation; the knowledge gained from our study could be further applied to other hepesviruses.
Animals ; Chick Embryo ; Chickens ; Chromosomes, Artificial, Bacterial ; DNA, Recombinant ; Herpesvirus 2, Gallid/genetics* ; Marek Disease

OBJECTIVE: To investigate the inhibitory effect of ketogenic diet (KD) on growth of neuroblastoma in mice. METHODS: BALB/c-nu mouse models bearing neuroblastoma xenografts were established by subcutaneous injection of human neuroblastoma cell line (SH-SY5Y). When the tumor volume reached 250 mm3, the mice were randomized into SD group with standard diet and PBS treatment, KD group with ketogenic diet and PBS treatment, and CP+KD group with ketogenic diet and cyclophosphamide (60 mg·kg·day) treatment, =8. The tumor volume, body weight, blood glucose, ketone body (β-Hydroxybutyrate) levels, and hepatic steatosis in the mice were assessed. The expressions of caspase-3 and caspase-8 were detected by Western blotting, and Ki67 expresison was detected using immunohistochemistry (IHC). Transmission electron microscopy (TEM) was employed for the autophagosomes, and the autophagic protein Beclin1, LC3A/B and P62 were detected by IHC and Western blotting. RESULTS: On day 28 post tumor cell injection, the mice in KD and CP+KD groups could prolong the overall survival rates than that in SD group ( < 0.001). On day 22 post the injection, the tumor volume in KD group was smaller than that in SD group ( < 0.05); on 16, 19, and 22 day post the injection, the tumor volume in CP+KD group was smaller than that in SD group ( < 0.01). The mice in SD group showed greater body weight on day 19 and higher blood glucose level on day 13 post the injection than those in the other two groups ( < 0.05). Blood ketone level and hepatic steatosis score were higher and glucose ketone index (GKI) was lower in KD and CP+KD groups than those in SD group (all < 0.05). The expressions of Ki67 and apoptotic proteins were detected in the tumor tissues of all groups. TEM revealed more autophagosomes in the tumor tissues of KD group than that of SD group. P62 expression was lowered ( < 0.01) and Beclin1 and LC3A/B expressions were up-regulated in the tumor tissues of KD group ( < 0.05), which is consisitent with IHC. CONCLUSIONS KD has a strong anti-tumor effect in the xenograft mouse model possibly by regulating cell autophagy.
3-Hydroxybutyric Acid ; Animals ; Blood Glucose ; Cell Line, Tumor ; Diet, Ketogenic ; Humans ; Mice ; Mice, Inbred BALB C ; Neuroblastoma

In the nucleus, chromatin is folded into hierarchical architecture that is tightly linked to various nuclear functions. However, the underlying molecular mechanisms that confer these archi-tectures remain incompletely understood. Here, we investigated the functional roles of H3 lysine 9 dimethylation (H3K9me2), one of the abundant histone modifications, in three-dimensional (3D) genome organization. Unlike in mouse embryonic stem cells, inhibition of methyltransferases G9a and GLP in differentiated cells eliminated H3K9me2 predominantly at A-type (active) genomic compartments, and the level of residual H3K9me2 modifications was strongly associated with B-type (inactive) genomic compartments. Furthermore, chemical inhibition of G9a/GLP in mouse hepatocytes led to decreased chromatin-nuclear lamina interactions mainly at G9a/GLP-sensitive regions, increased degree of genomic compartmentalization, and up-regulation of hundreds of genes that were associated with alterations of the 3D chromatin. Collectively, our data demonstrated essential roles of H3K9me2 in 3D genome organization.