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*

BACKGROUND: In China, lung cancer remains the cancer with the highest incidence and mortality rate. Among early-stage lung adenocarcinomas (LUAD), the micropapillary (MPP) component is prevalent and typically exhibits high aggressiveness, significantly correlating with early metastasis, lymphatic infiltration, and reduced five-year survival rates. Therefore, the study is to explore the similarities and differences between MPP and non-micropapillary (non-MPP) components in malignant pulmonary nodules characterized by GGOs in early-stage LUAD, identify unique mutational features of the MPP component and analyze the relationship between the ZNF469 gene, a member of the zinc-finger protein family, and the prognosis of early-stage LUAD, as well as its correlation with immune infiltration. METHODS: A total of 31 malignant pulmonary nodules of LUAD were collected and dissected into paired MPP and non-MPP components using microdissection. Whole-exome sequencing (WES) was performed on the components of early-stage malignant pulmonary nodules. Mutational signatures analysis was conducted using R packages such as maftools, Nonnegative Matrix Factorization (NMF), and Sigminer to unveil the genomic mutational characteristics unique to MPP components in invasive LUAD compared to other tumor tissues. Furthermore, we explored the expression of the ZNF469 gene in LUAD using The Cancer Genome Atlas (TCGA) database to investigate its potential association with the prognosis. We also investigated gene interaction networks and signaling pathways related to ZNF469 in LUAD using the GeneMANIA database and conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Lastly, we analyzed the correlation between ZNF469 gene expression and levels of immune cell infiltration in LUAD using the TIMER and TISIDB databases. RESULTS: MPP components exhibited a higher number of genomic variations, particularly the 13th COSMIC (Catalogue of Somatic Mutations in Cancer) mutational signature characterized by the activity of the cytidine deaminase APOBEC family, which was unique to MPP components compared to non-MPP components in tumor tissues. This suggests the potential involvement of APOBEC in the progression of MPP components in early-stage LUAD. Additionally, MPP samples with high similarity to APOBEC signature displayed a higher tumor mutational burden (TMB), indicating that these patients may be more likely to benefit from immunotherapy. The expression of ZNF469 was significantly upregulated in LUAD compared to normal tissue, and was associated with poor prognosis in LUAD patients (P<0.05). Gene interaction network analysis and GO/KEGG enrichment analysis revealed that COL6A1, COL1A1, COL1A2, TGFB2, MMP2, COL8A2 and C2CD4C interacted with ZNF469 and were mainly involved in encoding collagen proteins and participating in the constitution of extracellular matrix. ZNF469 expression was positively correlated with immune cell infiltration in LUAD (P<0.05). CONCLUSIONS The study has unveiled distinctive mutational signatures in the MPP components of early-stage invasive LUAD in the Asian population. Furthermore, we have identified that the elevated expression of mutated ZNF469 impacts the prognosis and immune infiltration in LUAD, suggesting its potential as a diagnostic and prognostic biomarker in LUAD.
Humans ; Lung Neoplasms/genetics* ; Adenocarcinoma of Lung/genetics* ; China ; Prognosis ; Transcription Factors

Objective To investigate the effects of long-term administration of tacrolimus(also known as FK506)on the pain-related behaviors in mice and to study the underlying mechanism of pain induced by FK506 via measuring the effect of FK506 on the synaptic expression and phosphorylation of alpha-amino-3-hyroxy-5-methyl-4-isoxazolepropionic acid(AMPA)receptor in the spinal cord dorsal horn of mice.Methods 1)A total of 24 mice were evenly and randomly assigned to two groups,a FK506 group and a Saline group.The FK506 group was given daily intraperitoneal injection of FK506 and the Saline group received normal saline.Both groups received injection once a day for 7 days in a row.Some of the mice(n=6 in each group)were monitored for the changes in the paw withdrawal threshold(PWT),the paw withdrawal latency(PWL),and the spontaneous pain behaviors to establish the pain model.The other mice(n=6 in each group)of each group underwent isolation of the dorsal horn when obvious pain symptoms were induced on day 7 of injection.Then,immunoblotting was performed to determine the synaptic expression and phosphorylation levels of GluA1 and GluA2 subunits of AMPA receptors.2)The mice were randomly divided into two groups,FK506+calcineurin(CaN)group and FK506+Saline group(n=6 in each group).After the pain model was constructed,the mice were given intrathecal injection of recombinant CaN(also know as 33 U)or normal saline.Then,60 minutes later,the PWT and the PWL of the mice were measured to investigate the role of CaN in FK506-induced pain.3)Another18 mice were selected.The mice were randomly and evenly assigned to three groups,a control group(receiving intraperitoneal injection of normal saline followed by intrathecal injection of normal saline),FK506+Saline group(receiving intraperitoneal injection of FK506 followed by intrathecal injection of normal saline)and FK506+CaN group(receiving intraperitoneal injection of FK506 followed by intrathecal injection of CaN).Then,60 minutes later,the spinal cords were isolated and subjected to immunoblotting assay to determine the role of CaN in FK506-induced AMPA receptor modification.Results 1)After 7 consecutive days of intraperitoneal injection of FK506,the PWT and PWL of mice dropped significantly,reaching on day 7 as low as 22.3%±0.05%and 66.6%±0.05%of the control group,respectively(P<0.01).The FK506-treated mice displayed evident spontaneous pain behavior,presenting significantly increased licking activities(P<0.01).These results indicated that FK506-induced pain model was successfully established.Immunoblotting assay showed that the total expressions of GluA1 and GluA2 subunits in the spinal dorsal horn of the FK506 group remained unchanged in comparison with those of the Saline group.However,FK506 specifically induced an increase in the synaptic expression of GluA1.In addition,the phosphorylation levels of GluA1 at Ser845 and Ser831 in FK506-treated mice were significantly increased in comparison with those of the control group(P<0.05).2)Compared with those of the mice in the FK506+Saline group,the PWT and the PWL of mice in the FK506+CaN group were significantly increased(P<0.05).3)Compared with those of the FK506+Saline group,the synaptic expression of GluA1 were decreased in FK506+CaN group(P<0.01)and the phosphorylation levels of GluA1 at Ser845 and Ser831 were significantly downregulated(P<0.001).Conclusion The hyper-expression and hyperphosphorylation of GluA1 subunit in the spinal cord dorsal horn resulting from CaN inhibition contributes to the FK506-induced pain syndrome.FK506 induces the synaptic hyper-expression and hyperphosphorylation of GluA1 in the dorsal horn of the spinal cord through CaN inhibition,thereby inducing pain.

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 compare the clinical effects of minimally invasive reduction through a bone tunnel combined with Jail screwing and those of posterolateral locking plating in the treatment of simple posterolateral tibial plateau fractures.Methods:A retrospective analysis was conducted of the data of 48 patients who had been operatively treated and completely followed up at Department of Orthopedics, Suqian Hospital of Nanjing Drum Tower Hospital Group for simple posterolateral tibial plateau fractures from October 2016 to October 2020. There were 26 males and 22 females, aged from 35 to 68 years. They were divided into a minimally invasive group (25 cases subjected to minimally invasive reduction through a bone tunnel combined with Jail screwing) and an incision group (23 cases subjected to posterolateral locking plating) according to their surgical methods. The operation time, incision length, intraoperative blood loss, fracture healing time, cumulative fluoroscopy time, hospital stay and posterior inclination angles of the tibial plateau and Hospital for Special Surgery (HSS) knee function scores at 1, 3, 6, 9, and 12 months after operation were compared between the 2 groups. Complications in the 2 groups of patients were recorded.Results:There was no significant difference in the preoperative general data between the 2 groups, showing comparability ( P>0.05). The 48 patients were followed up for 12 to 36 months (average 16.5 months). The minimally invasive group was significantly better than the incision group in operation time [(42.6±9.1) min versus (65.7±11.5) min], incision length [(4.0±0.4) cm versus (15.0±1.5) cm], intraoperative blood loss[(22.6±5.8) mL versus (31.5±8.8) mL], hospital stay [(7.6±1.4) d versus (11.1±2.4) d], and HSS score one month after operation [(84.8±1.9) points versus (72.9±4.1) points], but the cumulative fluoroscopy time in the incision group [(4.1±1.4) s]was significantly less than that in the minimally invasive group [(22.3±4.2) s] ( P<0.05). There were no significant differences in fracture healing time, HSS scores at 3, 6, 9, or 12 months after operation, or posterior inclination angle of the tibial plateau between the 2 groups ( P>0.05). There were no such complications as wound infection, vascular injury, internal fixation failure, nonunion or malunion of fractures in either of the 2 groups. Two cases in the incision group presented with symptoms of common peroneal nerve injury but recovered 3 months after operation. Conclusions:Although both minimally invasive reduction through a bone tunnel combined with Jail screwing and posterolateral locking plating can achieve satisfactory outcomes in the treatment of simple posterolateral tibial plateau fractures, the minimally invasive technique is preferable because it shows the advantages of a smaller incision, less bleeding, shorter operation time, a lower operation risk, quicker postoperative recovery and shorter hospital stay.

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.

"Internet + Nursing" can break the limits of time and space, not only provide high-quality nursing services conveniently and quickly, but also maximize limited medical care resources. "Internet + Nursing" has become the focus of the national medical and health care reform and the development direction of continuous nursing for chronic wounds. This article reviews the application concepts and methods of "Internet + Nursing" in chronic wound management, and so as to provide a reference for the further development of "Internet + Nursing" in chronic wound management.

Background and objective Cetuximab is a monoclonal antibody directed against epidermal growth fac-tor receptor. Emerging evidence showed improved effcacy with the addition of cetuximab to chemotherapy in advanced non-small cell lung cancer (NSCLC), but the data in oriental population are limited. hTe aim of this study is to investigate the eff-cacy of cetuximab in combination with chemotherapy in Chinese patients with advanced NSCLC.Methods NSCLC patients receiving cetuximab in combination with chemotherapy in Cancer Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College were enrolled and retrospectively analyzed. Clinical characteristic, effcacy, outcome and toxicity data were analyzed.Results A total of 40 patients were enrolled into this study in which 29 were male, 36 with adenocarcinoma. In the 23 patients who had received palliative chemotherapy previously (with a median of 2 prior chemotherapy regimens), the median progression-free survival (PFS) atfer the last prior chemotherapy regimen was 2.3 months. For the overall population, 13 (32.5%) patients achieved partial response atfer cetuximab in combination with chemotherapy. Response rate were 52.9% (9/17) and 17.4% (4/23) in chemotherapy-naive patients and chemotherapy-treated patients, respectively (P=0.018). hTe median PFS was 4.8 months for the overall population. In chemotherapy-naive patients and chemotherapy-treated patients, the median PFS was 8.4 months and 4.1 months, respectively (P=0.062). hTe estimated median overall survival was 17.1 months. Toxicities were generally manageable and no treatment-related deaths occurred.Conclusion Cetuximab in addition to che-motherapy appears to be associated with promising effcacy and acceptable toxicity proifle in Chinese patients with advanced NSCLC. Further validation is needed.

Background and objective Brain metastasis was frequent in non-small cell lung cancer (NSCLC) patients with poor prognosis. Geiftinib was an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor which has been used in the treatment of NSCLC. Our study was to evaluate the effcacy and toxicities of geiftinib in lung adenocarcinoma patients with brain metastases. Methods We retrospectively reviewed clinical records of 32 lung adenocarcinoma patients with brain metastases, who had received geiftinib 250 mg Qd until disease progression or intolerable toxicities. Results hTe median overall survival (mOS) and median progression-free survival (mPFS) were 24.7 months and 11.2 months, respectively. Response rate (RR) and disease control rate (DCR) were 62.5%and 93.8%, respectively. hTe mOS and mPFS of geiftinib-naive patients were 35.6 months and 11.3 months, respectively, and RR and DCR were 75.0%and 100.0%, respectively. hTe mOS and mPFS of geiftinib treatment patients were 18.6 months and 6.7 months, respectively, and RR and DCR were 50.0%and 83.3%, respectively. hTe mOS and mPFS of patients with sensitive EGFR mutation were 24.8 months and 10.8 months, respectively, and RR and DCR were 75.0%and 100.0%, respectively. hTe mOS and mPFS of patients with unknown EGFR status were 35.6 months and 12.3 months, respectively, and RR and DCR were 53.3%and 86.7%, respectively. Treatment was well tolerated and no severe toxicities were observed. Common toxicities include:rash in 15 patients (46.9%), diarrhea in 7 cases (21.9%) and oral ulcer in 1 case (3.1%). Conclusion Geiftinib was highly effective and well tolerated in lung adenocarcinoma patients with brain metastases, and could be recommended as a treatment choice for this population.

Aim To explore new ways for developing anticancer drugs by the separation of pigment from Fu-sarium species JN158 ( Fusarium sp JN158 ) , the iden-tification of its structure, the screening of anticancer components and the study of its partial mechanism. Methods Pigment separation was done by HPLC, structural analysis by UV, IR, NMR, the screening of anticancer activity by MTT. Western blot was used to analyze the protein expression of CyclinD1, NF-κB, VEGF in tumor cells. Results The results showed that the pigment from Fusarium produced a total of six different peaks, of which peak Ⅵ was the anthocya-nins. Its molecular weight is about 382, molecular for-mula is C17 H18 O10 . According to investigation, this pig-ment was probably a new compound, which could in-hibit the proliferation of MCF-7 cells markedly ( IC50:0.011mmol·L-1 ,P<0.05;the control medicine ube-nimex IC50:10 mmol · L-1 ) in a concentration-de-pendent manner, and had no effect on human umbilical cord intravenous endotheliocyte ( HUVEC ) . The influ-ence on the gene expression of CyclinD1, NF-κB, VEGF in MCF-7 cells varied with the concentration of this compound. The Western blot results showed that VI pigment compound inhibited CyclinD1, NF-κB, VEGF gene expression (P<0.05 or 0. 01),compared with the control group. Conclusion The Ⅵ pigment compound from Fusarium sp JN158 could inhibit MCF-7 proliferation by inhibiting CyclinD1, NF-κB, VEGF gene expression. The compound may be a promising compound against breast cancer.