Biological activity of recombinant bovine IFN-α and inhibitory effect on BVDV in vitro.

Type I interferon has great broad-spectrum antiviral ability and immunomodulatory function, and its receptors are expressed in almost all types of cells. Bovine viral diarrhea virus (BVDV) is an important pathogen causing significant economic losses in cattle. In this study, a recombinant expression plasmid carrying bovine interferon-α(BoIFN-α)gene was constructed and transformed into E. coli BL21 (DE3) competent cells. SDS-PAGE and Westernblotting analysis showed that the recombinant BoIFN-α protein (rBoIFN-α) was successfully expressed. It is about 36KD and exists in the form of inclusion body. When denatured, purified and renatured rBoIFN-α protein stimulated MDBK cells, the expression of interferon stimulating genes (ISGs) such as ISG15, OAS1, IFIT1, Mx1 and IFITM1 were significantly up-regulated, and reached the peak at 12 h (P< 0.001). MDBK cells were infected with BVDV with moi of 0.1 and 1.0, respectively. The virus proliferation was observed after pretreatment with rBoIFN-α protein and post-infection treatment. The results showed that the denatured, purified and renatured BoIFN-α protein had good biological activity and could inhibit the replication of BVDV in MDBK cells in vitro, which provided a basis for BoIFN-α as an antiviral drug, immune enhancer and clinical application of BVDV.

High expression of centromere protein N as novel biomarkers for gastric adenocarcinoma.

BACKGROUND: The role and mechanism of centromeric protein N (CENP-N), which has been associated with the development of various cancer types, are yet unclear in stomach adenocarcinoma (STAD). METHODS: Data from the Cancer Genome Atlas and Genotype-Tissue Expression were used to determine whether CENP-N expression was altered in STAD tumors compared to normal tissues. Xiantao was used to perform Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis on CENP-N. The relationship between CENP-N expression and immune cell infiltration was assessed using TCGA database. The expression of CENP-N in STAD and surrounding tissues was confirmed using immunohistochemical staining and the correlation between CENP-N expression and clinicopathological characteristics was examined. The effects of CENP-N knockdown by siRNA on proliferation were measured by CCK-8 and EdU assays in AGS cells. Following siRNA transfection, flow cytometry was performed to evaluate cell cycle and apoptotic alterations in AGS cells. The effect of CENP-N knockdown on the expression level of related proteins was detected by Westren blot. RESULTS: CENP-N was highly expressed in STAD tissues, which was confirmed by our immunohistochemistry results. The degree of invasion, TNM stage, and lymph node metastases were all strongly associated with CENP-N expression. CENP-N was essential for the cell cycle, DNA replication, chromosomal segregation, and nuclear division; there was a positive correlation between CENP-N expression and infiltrating Th2 and NK CD56dim cells and a negative correlation between CENP-N expression and mast, pDC, NK, and B cell infiltration. When CENP-N expression in AGS cells was knocked down, cell proliferation dramatically reduced (p < .05) and the percentage of cells in the S and G2-M phases decreased significantly (p < .05). Silencing CENP-N significantly promoted the apoptosis of AGS cells (p < .05). Mechanistic investigations showed that silencing CENP-N expression may inhibit STAD proliferation through the Cyclin E1 and promote STAD apoptosis through the Bcl-2/Bax. CONCLUSION: According to our data, CENP-N acts as an oncogene in STAD and may be a viable therapeutic target.

Linguistic indicators for predicting the veracity of online health rumors.

This study aims to examine the role of language in discerning the authenticity of online health rumors. To achieve this goal, it specifically focuses on analyzing five categories of linguistic indicators: (1) emotional language characterized by sentiment words, sensory words, and continuous punctuations, (2) exaggerated language defined by the presence of extreme numbers and extreme adverbs, (3) personalized language denoted by first-person pronouns, (4) unprofessional language represented by typographical errors, and (5) linkage language marked by inclusion of hyperlinks. To conduct the investigation, a dataset consisting of 1,500 information items was utilized. The dataset exhibited a distribution pattern wherein 20% of the information was verified to be true, while the remaining 80% was categorized as rumors. These items were sourced from two prominent rumor-clarification websites in China. A binomial logistic regression was used for data analysis to determine whether the language used in an online health rumor could predict its authenticity. The results of the analysis showed that the presence of sentiment words, continuous punctuation marks, extreme numbers and adverbs in an online health rumor could predict its authenticity. Personalized language, typographical errors, and hyperlinks were also found to be useful indicators for identifying health rumors using linguistic indicators. These results provide valuable insights for identifying health rumors using language-based features and could help individuals and organizations better understand the credibility of online health information.

Prognostic analysis of patients with combined hepatocellular-cholangiocarcinoma after radical resection: A retrospective multicenter cohort study.

BACKGROUND: Combined hepatocellular-cholangiocarcinoma (cHCC-CCA) is a form of rare primary liver cancer that combines intrahepatic cholangiocarcinoma (ICC) and hepatocellular carcinoma. AIM: To investigate overall survival (OS) and recurrence-free survival (RFS) after radical resection in patients with cHCC-CCA, and the clinicopathological factors affecting prognosis in two center hospitals of China. METHODS: We reviewed consecutive patients with cHCC-CCA who received radical resection between January 2005 and September 2021 at Peking Union Medical College and the 5th Medical Center of the PLA General Hospital retrospectively. Regular follow-up and clinicopathological characteristics were systematic collected for baseline and prognostic analysis. RESULTS: Our study included 95 patients who received radical resection. The majority of these patients were male and 82.7% of these patients were infected with HBV. The mean tumor size was 4.5 cm, and approximately 40% of patients had more than one lesion. The median OS was 26.8 (95%CI: 18.5-43.0) mo, and the median RFS was 7.27 (95%CI: 5.83-10.3) mo. Independent predictors of OS were CA19-9 ≥ 37 U/mL (HR = 8.68, P = 0.002), Child-Pugh score > 5 (HR = 5.52, P = 0.027), tumor number > 1 (HR = 30.85, P = 0.002), tumor size and transarterial chemoembolization (TACE) after surgery (HR = 0.2, P = 0.005). CONCLUSION: The overall postoperative survival of cHCC-CCA patients is poor, and most patients experience relapse within a short period of time after surgery. Preoperative tumor biomarker (CA19-9, alpha-fetoprotein) levels, tumor size, and Child-Pugh score can significantly affect OS. Adjuvant TACE after surgery prolongs RFS, suggesting that TACE is a possible option for postoperative adjuvant therapy in patients with cHCC-CCA.

Age, gender, geographic and clinical differences for gallstones in China: a nationwide study.

Background: The etiology of gallstones at different anatomical locations may be varied. However, it has not been well studied about the prevalence of gallstones by anatomical locations and their associated factors in China. Methods: We used the data set from preventive health screening centers covering all provinces in mainland China except for Tibet, and a total of 10,937,993 adults were included, who received abdominal ultrasonography in 2017. We analyzed the prevalence of gallstones classified by anatomical locations, including gallbladder (GB) stones, intrahepatic bile duct (IHD) stones, and extrahepatic bile duct (EHD) stones. Further, their associated factors were investigated using a logistic regression model with body mass index (BMI), fasting glucose, total triglyceride, and previous cholecystectomy, with covariates of age and sex. Results: The age- and gender-standardized prevalence (AGS-prevalence) of gallstone diseases was 5.13% (95% CI: 5.11-5.14%). GB, IHD, and EHD stones accounted for 76.3%, 24.3%, and 0.2% of all gallstone cases (concomitant cases were present). GB, IHD, and EHD gallstones presented different patterns by the age, gender, geographic and metabolic factors. Overall, the age-standardized prevalence was higher in women than that in men (5.41% vs. 4.85%, P<0.001). The gender standardized prevalence of all gallstone subtypes apparently increased with age (P<0.001), especially for GB stones from 1.05% (age 18-30) to 11.6% (age ≥70) (P<0.001). There was a marked geographic variation with AGS-prevalence ranging from 3.00% to 8.86% among different provinces. Noticeably, higher BMI, fasting glucose level, or total triglyceride level was associated with a higher prevalence of overall gallstones and GB stones (OR >1), but associated with a lower prevalence of IHD and EHD stones. Conclusions: The prevalence of gallstones in China largely varied in its anatomical location, demographic factors, geographic location, and metabolic factors, suggesting that the etiology of each subtype may be different. Further investigation should be conducted.

Content characteristics predict the putative authenticity of COVID-19 rumors.

Rumors regarding COVID-19 have been prevalent on the Internet and affect the control of the COVID-19 pandemic. Using 1,296 COVID-19 rumors collected from an online platform (piyao.org.cn) in China, we found measurable differences in the content characteristics between true and false rumors. We revealed that the length of a rumor's headline is negatively related to the probability of a rumor being true [odds ratio (OR) = 0.37, 95% CI (0.30, 0.44)]. In contrast, the length of a rumor's statement is positively related to this probability [OR = 1.11, 95% CI (1.09, 1.13)]. In addition, we found that a rumor is more likely to be true if it contains concrete places [OR = 20.83, 95% CI (9.60, 48.98)] and it specifies the date or time of events [OR = 22.31, 95% CI (9.63, 57.92)]. The rumor is also likely to be true when it does not evoke positive or negative emotions [OR = 0.15, 95% CI (0.08, 0.29)] and does not include a call for action [OR = 0.06, 95% CI (0.02, 0.12)]. By contrast, the presence of source cues [OR = 0.64, 95% CI (0.31, 1.28)] and visuals [OR = 1.41, 95% CI (0.53, 3.73)] is related to this probability with limited significance. Our findings provide some clues for identifying COVID-19 rumors using their content characteristics.

Molecular characterization of a novel subgenotype of lumpy skin disease virus strain isolated in Inner Mongolia of China.

BACKGROUND: The outbreak of Lumpy skin disease (LSD) in cattle caused by LSD virus (LSDV) was first reported in August 2019 in China. Since then, several LSD outbreaks have been reported in seven different provinces of China. Until now, several Lumpy skin disease virus (LSDV) strains from China have been reported and sequenced including LSDV/Xinjiang/2019 (MN598005.1), China/GD01/2020 (MW355944.1), and LSDV/Hongkong/2021 (MW732649.1). In October 2020, more than 1,700 cattle imported from Chile arrived in Xilingol, Inner Mongolia, and were diagnosed with LSD. Currently, limited data on the origin of the virus is available. METHODS: Nucleotide sequences of the ORF11, ORF36, ORF74, ORF117, ORF126 genes and the complete genome of LSDV strains and isolates were downloaded from NCBI database. MEGA7.0 was used to perform phylogenetic analysis with Neighbor-Joining (NJ). DNASTAR software is used to analyze homologous comparison analysis with related genes of reference strains included in Genbank. RESULTS: Compared with other strains isolated from China, the results of full genome sequence analysis showed the LSDV/NMG/2020 strain belonged to the recombinant strains. The LSDV/NMG/2020 strain is different from the current LSDV field isolates in Africa, the Middle East, Europe, and the newly emerged LSDV Russia variants. Based on the identities of P32, RPO30, EEV, GPCR and LSDV117 genes (99.8%, 99%, 99.8%, 99% and 98.7%), the sub-cluster recombinant containing LSDV/NMG/2020 strain is phylogenetically closer to the Russia strain (Saratov/2017). CONCLUSIONS: In this study, we reported a new isolated LSDV strain named LSDV/NMG/2020. The results of genomic characterization and phylogenetic analysis demonstrated that the LSDV/NMG/2020 isolate was a vaccine-like recombinant strain.

Phylogenetic and pathogenicity analysis of a novel lineage of caprine parainfluenza virus type 3.

Caprine parainfluenza virus type 3 (CPIV3) was first identified in goats named JS2013 in China. In 2019, a sheep herd broke a disease with respiratory disease in Hebei province, China. In order to confirm the pathogen of the disease, the nasal swabs, stool swabs and blood samples were collected from the sheep. Virus isolation was performed on MDBK cells and identification was conducted by RT-PCR. The complete genome of the isolate was sequenced and phylogenetic analyzed. In order to evaluate the pathogenicity of the virus, five seronegative sheep were experimental infected with the virus suspension. The phylogenetic analyses based on the complete genome and the M gene indicated that the isolate strain was distinguished distinct from previously reported CPIV3 lineage of JS2013. The virus-inoculated sheep displayed the syndrome with depression, cough, and fever. Virus shedding were detected by RT-PCR from nasal swabs. All infected showed virus shedding during 2 - 21dpi and viremia could be detected in serum samples. Gross pathological assessment of sheep in infected group showed gross lesion in the lungs. Histopathological observation results indicated that lungs had mild to moderate interstitial pneumonia, with thickened alveolar walls, decreased alveolar space, and increased amounts of inflammatory cells infiltration. This is the first report of pathogenicity of the novel lineage of sheep-derived CPIV3. The results would be helpful for further studies on the prevention and control strategies for CPIV3 infections in goat and sheep.

Hsa_circ_101882 promotes migration and invasion of gastric cancer cells by regulating EMT.

BACKGROUND: At present, gastric cancer (GC) is a serious threat to human life and health. Non-coding circular RNAs (circRNAs) have been found abnormal expression in multiple tumors. However, circRNAs remain largely unknown in tumor progression. In the present study, we mainly examined the expression, function, and molecular mechanisms of a new circRNAs (hsa_circ_101882) in GC. MATERIALS AND METHODS: The expression of hsa_circ_101882 in GC tissue, corresponding adjacent normal tissues, and GC cell lines was examined by RT-PCR. The function of hsa_circ_101882 in GC was evaluated by MTT assay, cell migration, and invasion assay, colony formation assay, and flow cytometric assay. The effect of hsa_circ_101882 on epithelial-to-mesenchymal transition (EMT)-related gene expression was detected by RT-PCR and Western blot. RESULTS: Hsa_circ_101882 expression levels were significantly increased in GC tissue and GC cell lines. Functionally, low expression of hsa_circ_101882 revealed anti-tumor effects via inhibiting cell growth, migration, and invasion and promoting cell apoptosis. Mechanically, the dysregulated expression of hsa_circ_101882 affects EMT signaling pathway, which was examined by detecting E-cadherin, N-cadherin, vimentin, and Snail expression levels. CONCLUSIONS: Therefore, our research reveals that hsa_circ_101882 is considered a metastasis promoter by activating EMT and may serve as a critical oncogene and potential new biomarker in GC.

NIR absorbing reduced graphene oxide for photothermal radiotherapy for treatment of esophageal cancer.

Theranostic agents were drawing a huge attention in the personalized medication. In this study, we established a facile technique, plant extract-based technique for the synthesis of reduced nano-graphene oxide (RNGO) with low cytotoxicity. We formed platforms of photothermal (PT) therapy and further explained that the synthesized RNGO can be utilized as ready to use PT therapy without any additional surface adjustment. In the meantime, with the help of a constant-wave NIR laser (near-infrared), in vitro esophageal adenocarcinoma cell line (OE-19) cancer cells were effectively ablated, because of the PT impact of RNGO. The outcomes propose that the RNGO was appropriate for PT therapy and photoacoustic imaging of the tumor, which is assuring for theranostic nanomedicine.

Hepatobiliary case report and literature review of hepatic reactive lymphoid hyperplasia with positive anti-smooth muscle antibody and anti-nuclear antibody tests.

Hepatic reactive lymphoid hyperplasia (RLH) is a rare benign tumor of the liver that exhibits similar imaging characteristics to those of other hepatic malignant tumors; therefore, it requires novel biomarkers to be differentiated from the other tumors. A 69-year-old female was found to have a mass in the liver and was admitted to Peking Union Medical College Hospital for further evaluation. Most laboratory tests, including liver function tests, tumor biomarkers, and autoimmune markers were within normal range, except for positive antinuclear antibody and anti-smooth muscle antibody (SMA) tests. Diagnostic imaging, including ultrasonography, computerized tomography and magnetic resonance imaging displayed a small hepatic mass suggestive of hepatocellular carcinoma (HCC). Partial hepatectomy was performed, and histological diagnosis suggested RLH. Postoperative treatments included anti-infection, nutritional enhancement and liver protection. The suspicion of autoimmune hepatitis could not be confirmed. No recurrence or autoimmune disease was observed over 6-month follow-up. Positive anti-nuclear antibody (ANA) and anti-SMA may be potential biomarkers for hepatic RLH.

Cell-free plasma hypermethylated CASZ1, CDH13 and ING2 are promising biomarkers of esophageal cancer.

Identifying sensitive and specific biomarkers for early detection of cancer is immensely imperative for early diagnosis and treatment and better clinical outcome of cancer patients. This study aimed to construct a specific DNA methylation pattern of cancer suppressor genes and explore the feasibility of applying cell-free DNA based methylation as a biomarker for early diagnosis of esophageal squamous cell carcinoma (ESCC). We recruited early stage ESCC patients from Yangzhong County, China. The Illumina Infinium 450K Methylation BeadChip was used to construct a genome-wide DNA methylation profile. Then, differentiated genes were selected for the validation study using the Sequenom MassARRAY platform. The frequency of methylation was compared between cancer tissues, matched cell-free DNAs and normal controls. The specific methylation profiles were constructed, and the sensitivity and specificity were calculated. Seven CG sites in three genes CASZ1, CDH13 and ING2 were significantly hypermethylated in ESCC as compared with normal controls. A significant correlation was found between the methylation of DNA extracted from cancer tissues and matched plasma cell-free DNA, either for individual CG site or for cumulative methylation analysis. The sensitivity and specificity reached 100% at an appropriate cut-point using these specific methylation biomarkers. This study revealed that aberrant DNA methylation is a promising biomarker for molecular diagnosis of esophageal cancer. Hypermethylation of CASZ1, CDH13 and ING2 detected in plasma cell-free DNA can be applied as a potential noninvasive biomarker for diagnosis of esophageal cancer.

[A real-time fluorescence quantitative PCR method for source identification of Crocus sativus and Carthamus tinctorius].

The specific PCR primer was designed base on ITS2 sequence in GenBank, and we developed a SYBRGreen real-time fluorescence quantitative PCR system for identification of Crocus sativus and Carthamus tinctorius source. Compared with Chinese herbal medicine DNA barcode technique, this method showed characteristics of shorter time, higher specificity and sensitivity. Using this method to detect 15 samples, 4 were C. sativus, 8 were C. tinctorius, and the other 3 samples were none of them. The result was in accordance with Chinese herbal medicine DNA barcode. This study lays the foundation for identification of related Chinese medical materials.

Lentivirus-mediated RNA interference targeting UbcH10 reduces cell growth and invasion of human osteosarcoma cells via inhibition of Ki-67 and matrix metalloproteinases.

Osteosarcoma (OS) is the most commonly diagnosed primary malignancy affecting the bone. UbcH10 is a cancer-related E2-ubiquitin-conjugating enzyme. An overexpression of UbcH10 is significantly associated with tumor grade and cellular proliferation. However, limited evidence exists with regard to the biological function of UbcH10 in OS. The present study created a UbcH10 knockdown OS cell line using lentivirus-mediated RNA interference. The expression of UbcH10 was significantly reduced in UbcH10-targeted small hairpin RNA-expressing lentivirus OS cells. The downregulation of UbcH10 suppressed OS cell proliferation and colony formation ability via decreased Ki-67 expression. UbcH10 knockdown OS cells exhibited impaired invasion and migration abilities. Furthermore, knockdown of UbcH10 led to decreased levels of matrix metalloproteinase-3 and -9 in OS cells. The present study demonstrated the role of UbcH10 in OS cell proliferation, invasion and migration, which suggests that UbcH10 may be a potential candidate for OS therapy.

New fracture pattern focusing on implant fracture for periprosthetic femoral fractures.

PURPOSE: The Vancouver Classification System (VCS) for assessing periprosthetic femoral fractures has become universally accepted. The Unified Classification System (UCS) has expanded upon and updated the VCS and applied treatment principles to all periprosthetic fractures. However, periprosthetic femoral fractures accompanied by stem fracture after hip arthroplasty were not classifiable under the original VCS or the UCS. RESULTS: Our new fracture pattern is based on the periprosthetic femoral fracture as well as stem fracture after hip arthroplasty, and its treatment is dependent upon the stability of the proximal portion of the fractured femoral prosthesis. CONCLUSION: We believe that our new fracture pattern, a supplement to the VCS and UCS, is useful in the establishment of a therapeutic strategy for periprosthetic femoral fractures.

Co and Cr accumulation in hair after metal-on-metal hip resurfacing arthroplasty.

BACKGROUND: Currently, it is unclear whether Co and Cr levels are persistently elevated in the body after hip resurfacing arthroplasty (HRA). This study aimed to evaluate Co and Cr levels in the body after HRA using hair as samples. METHODS: Among 22 patients who had metal-on-metal HRA(m-o-m group) using the DePuy ASR system, scalp hair of 1.5 cm in length and 2 g in weight was collected before and at 6 and 12 months after surgery. Inductively coupled plasma-mass spectrometry (ICP-MS) was used to detect Co and Cr levels in the scalp hair. Another 22 patients with total hip replacements were randomly selected from a similar age range as controls (m-o-p group). Metal levels in hair were detected with multiple measurements and analysed by repeated measures analysis of variance. RESULTS: Both Co and Cr levels were significantly increased in the m-o-m group after surgery compared with those in the m-o-p group (P < 0.01). Co levels at 6 months post-operatively were 12-fold those before surgery, and Co levels 12 months postoperatively were lower than those at 6 months, but this difference was not significant (P = 0.0805) between the two time points in the m-o-m group. Cr levels were persistently elevated after surgery and were increased by 10-fold compared with those at baseline in the m-o-m group. CONCLUSIONS: Co and Cr levels in scalp hair are ideal indicators for metal levels in the body after HRA. Co and Cr levels in the body were greatly increased after metal-on-metal HRA. Long-term accumulation of these metals needs to be further investigated

A sustainable landscape ecosystem design: a case study.

Landscape planning is clearly ecologically and socially relevant. Concern about sustainability between human and environment is now a driving paradigm for this professional. However, the explosion of the sustainable landscape in China is a very recent phenomenon. What is the sustainable landscape? How is this realized in practice? In this article, on the basis of the reviews of history and perplexities of Chinese landscape and nature analysis of sustainable landscape, the ecothinking model, an implemental tool for sustainable landscape, was developed, which applies ecothinking in vision, culture, conservation and development of site, and the process of public participation for a harmonious relationship between human and environment. And a case study of the south entrance of TongNiuling Scenic Area was carried out, in which the most optimum scenario was chosen from among three models according to the ecothinking model, to illustrate the construction of the ecothinking model and how to achieve a sustainable landscape.

[Study on relationships between seven microsatellite loci and somatic cell score in Beijing Holstein cows].

Genetic variation of seven microsatellite loci BM1818, BM1258, BM1443, BM1905, BM302, BM4505 and CYP21 which were closely linked to somatic cell score (SCS) was analyzed in 240 Beijing Holstein cows with nondenaturing polyacrylamide gel electrophoresis. Allele frequencies, heterozygosity, polymorphic information content, the effective number of alleles of seven microsatellite loci were calculated. Relationships between seven microsatellite loci and somatic cell score in Beijing Holstein cows were primarily analyzed by least squares linear model. Least squares means of SCS for BM1818 (284 bp/284 bp), BM1258 (106 bp/92 bp), BM1443 (166 bp/160 bp), BM1905 (187 bp/187 bp), BM302 (142 bp/140 bp), BM4505 (240 bp/236 bp) and CYP21 (215 bp/198 bp) were relatively lower,and their genotypes were the most favorable genotypes in respective locus for mastitis resistance. Least squares means of SCS for BM1818 (286 bp/286 bp), BM1258 (102 bp/102 bp), BM1443 (170 bp/160 bp), BM1905 (197 bp/195 bp), BM302 (154 bp/145 bp), BM4505 (240 bp/238 bp) and CYP21 (204 bp/192 bp) were relatively higher,and their genotypes were the most unfavorable genotypes in respective locus for mastitis resistance. The information found in the present study would be very important for improving mastitis resistance in dairy cattle by marker assisted selection.