Quantitative analysis of microglia morphological changes in the hypothalamus of chronically stressed rats.

Based on the successful establishment of a rat model of chronic restraint stress, we used multiple algorithms to quantify the morphological changes of rat hypothalamic microglia from various perspectives, providing a pathomorphological basis for the subsequent study of molecular mechanisms of hypothalamic stress injury, such as neuroinflammation. To verify the successful establishment of the chronic stress model, an enzyme-linked immunosorbent assay was performed to detect serum glucocorticoid levels. Microglia labeled with Iba1 in frozen sections of rat hypothalamus were scanned and photographed at multiple levels using confocal microscopy. Subsequently, images were processed for external contouring and skeletonization, and morphological indices of microglia were calculated and analyzed using fractal, skeleton, and Sholl analysis. In addition, the co-expression of CD68 (a marker that can reflect phagocytic activity) and Iba1 was observed by immunofluorescence technique. Compared with the control group, microglia in the chronic stress group displayed reduced fractal dimension and lacunarity, increased density and circularity, enlarged soma areas, and shortened and reduced branches. Sholl analysis confirmed the reduced complexity of microglia following chronic stress. Meanwhile, microglia CD68 increased significantly, indicating that the microglia in the chronic stress group have greater phagocytosis activity. In summary, chronic restraint stress promoted the conversion of microglia in the rat hypothalamus to a less complex form, manifested as larger soma, shorter and fewer branches, more uniform and dense texture, and increased circularity; indeed, the shape of these microglia resembled that of amoeba and they displayed strong phagocytosis activity.

[Determination of 14 paralytic shellfish toxins in plasma and urine by ultra-high performance liquid chromatography-tandem mass spectrometry].

The detection of paralytic shellfish toxins in human biological matrices is important for the diagnosis and treatment of food poisoning caused by them. An ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for the determination of 14 paralytic shellfish toxins in plasma and urine. The effect of solid phase extraction (SPE) cartridges was also investigated and the pretreatment and chromatographic conditions were optimized. Under these optimal conditions, 0.2 mL water, 0.4 mL methanol, and 0.6 mL acetonitrile were successively added to plasma and urine samples for extraction. The supernatants from plasma extraction were subjected to an UHPLC-MS/MS analysis, whereas the supernatants from urine extraction were further purified using polyamide (PA) SPE cartridges and then analyzed by UHPLC-MS/MS. Chromatographic separation was conducted on a Poroshell 120 HILIC-Z column (100 mm×2.1 mm, 2.7 µm) with a flow rate of 0.5 mL/min. The mobile phase was 0.1% (v/v) formic acid aqueous solution containing 5 mmoL/L ammonium formate and acetonitrile containing 0.1% (v/v) formic acid. The analytes were detected in the multiple reaction monitoring (MRM) mode after being ionized by an electrospray ion (ESI) in positive and negative modes. Quantitation of the target compounds was performed using the external standard method. Under the optimal conditions, the method showed good linearity in the range of 0.24-84.06 µg/L, with correlation coefficients greater than 0.995. The limits of quantification (LOQs) for the plasma and urine samples were 1.68-12.04 ng/mL and 4.80-34.4 ng/mL, respectively. The average recoveries for all the compounds were 70.4%-123.4% at spiked levels of 1, 2, and 10 times the LOQs, the intra-day precisions were 2.3%-19.1% and the inter-day precisions were 5.0%-16.0%. The established method was used to determine the target compounds in the plasma and urine from mice intraperitoneally injected with 14 shellfish toxins. All 14 toxins were detected in the 20 urine and 20 plasma samples, with contents of 19.40-55.60 µg/L and 8.75-13.86 µg/L, respectively. The method is simple, sensitive, and only requires a small amount of sample. Therefore, it is highly suitable for the rapid detection of paralytic shellfish toxins in plasma and urine.

ERH Gene and Its Role in Cancer Cells.

Cancer is a major public health problem worldwide. Studies on oncogenes and tumor-targeted therapies have become an important part of cancer treatment development. In this review, we summarize and systematically introduce the gene enhancer of rudimentary homolog (ERH), which encodes a highly conserved small molecule protein. ERH mainly exists as a protein partner in human cells. It is involved in pyrimidine metabolism and protein complexes, acts as a transcriptional repressor, and participates in cell cycle regulation. Moreover, it is involved in DNA damage repair, mRNA splicing, the process of microRNA hairpins as well as erythroid differentiation. There are many related studies on the role of ERH in cancer cells; however, there are none on tumor-targeted therapeutic drugs or related therapies based on the expression of ERH. This study will provide possible directions for oncologists to further their research studies in this field.

Esthetic proportions of maxillary anterior tooth among 300 normal occlusion college students in Shanxi province / 中华口腔医学杂志

Objective: To explore the relationship between the width ratios of maxillary anterior teeth, the width/height ratios of maxillary central incisor and the esthetic proportions among individual normal occlusion, and to provide reference for the esthetic design of anterior teeth. Methods: A total of 300 Shanxi Medical University students who were residents from Shanxi province with normal occlusion (110 males, 190 females, aged 18-30 years) were recruited in this study from October 2020 to March 2021. Standardized digital photographs of the maxillary anterior tooth in natural head position were obtained, the perceived width of the maxillary anterior teeth (maxillary central incisors, lateral incisors and canines) on the left and right sides and height of maxillary central incisor were measured on the standardized digital photographs, then the width ratios of adjacent maxillary anterior teeth including the lateral incisor/central incisor width ratio (LI∶CI), the canine/lateral incisor width ratio (C∶LI), and the width/height ratios (W/H ratio) of the maxillary central incisor were calculated. The independent sample t test was used to compare the sex differences of each measurement project, Single sample t test was used to compare the difference of adjacent maxillary anterior teeth width ratios with golden proportion (0.618 and 0.618), Preston proportion (0.66 and 0.84), the recurring esthetic dental (RED) proportion (0.70 and 0.70). The percentage of anterior tooth width/intercanine width was calculated, the number and proportion of teeth conforming to golden percentage (25%, 15%, 10%)±1% or modified golden percentage (22.5%, 15.0%, 12.5%)±1% were calculated. The number and proportion of width/height ratio of maxillary central incisor ranged from 0.75 to 0.85 was counted. Results: The widths of maxillary central incisor, lateral incisor and canine were (8.50±0.52), (6.23±0.53) and (5.18±0.55) mm, respectively, the corresponding tooth of male [(8.74±0.49), (6.37±0.52), (5.41±0.47) mm] was significantly higher than that of female [(8.37±0.50), (6.15±0.52), (5.04±0.54) mm] (t=6.40, 3.55, 6.23,P<0.05). The width ratio of maxillary lateral incisor/central incisor was 0.73±0.05, and there was no significant difference between genders (t=-1.06, P>0.05). The width ratio of canine/lateral incisor was 0.84±0.10, and it was significantly higher in male (0.85±0.10) than in female (0.82±0.10) (t=2.42, P<0.05). Two width ratios of maxillary anterior teeth were significantly different from golden proportion and the RED proportion (t=38.50, 35.74, 11.48, 22.20, P<0.05). The lateral incisor/central incisor was significantly different from that of Preston proportion (t=24.66, P<0.05), while the canine/lateral incisor was not significantly different from that of Preston proportion (t=-0.92, P>0.05). In this study, a total of 0% (0/600) of central incisors, 63.0% (378/600) of lateral incisors and 5.8% (35/600) of canines met the golden percentage±1%. There were 42.8% (257/600) of central incisors, 63.0% (378/600) of lateral incisors and 56.7% (340/600) of canines met the modified golden percentage±1%. The width/height ratio of maxillary central incisors was 0.86±0.08, and there was no significant difference between genders (t=-0.88, P>0.05). Only 36.3% (218/600) of the subjects in this study ranged from 0.75 to 0.85. Conclusions: Gender differences should be considered in the esthetic design of anterior teeth; for the width ratios of maxillary anterior teeth, the golden proportion, the RED proportion and golden percentage do not accord with the natural tooth morphological characteristics of Shanxi nationality college students in normal occlusion. Preston proportion and modified golden percentage are of more reference value. The width/height ratio of maxillary central incisors is different from 0.75-0.85.

Study of mass spectrometry imaging metabolomics of paclitaxel based on esophageal cancer multicellular tumor spheroids cultured in vitro / 药学学报

Multicellular tumor spheroids (MCTS) can simulate the structure and metabolic characteristics of tumors in vivo, which is of great significance to study the metabolic phenotype of tumor cells and the mechanism of drug intervention. In this study, esophageal cancer MCTS were constructed, and MCTS frozen sections were prepared after treated with different formulations of paclitaxel (PTX) including common PTX injection, PTX liposome and albumin bound PTX. MCTS mass spectrometry imaging analysis method was established by using air flow assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI). The visualization of the permeation and enrichment process of PTX in MCTs after PTX treatment was realized, and the spatially resolved metabolomics of PTX injection group was studied. The results showed that the permeation and enrichment behavior of PTX in MCTs model were related to the formulations. The changes of endogenous metabolites in MCTs of esophageal cancer after treated with PTX injection had temporal and spatial characteristics. The metabolic changes of MCTS during the initial 0-4 hours were dominated by the down-regulation of middle-high polarity metabolites and some lipids in the central region of MCTS, while the metabolic changes of MCTS during 8-72 hours were mainly up-regulated by lipid metabolites in the peripheral region of MCTS. The combination of in vivo tumor-associated MCTs model with label free, highly sensitive and high coverage mass spectrometry imaging technology provided a new method and strategy for the study of pharmacometabolomics.

Diabetes onset at an earlier age and high HbA1c levels as risk factors of diabetic retinopathy.

AIM: To assess the effect of age at diabetes onset and uncontrollable high HbA1c levels on the development of diabetic retinopathy (DR) among Chinese type 2 diabetes mellitus (DM) patients. METHODS: This was a cross-sectional survey of diabetic patients in Subei district, China. Data covering physical measurements, fasting blood-glucose (FBG), glycosylated hemoglobin (HbA1c), blood lipid, urinary albumin/creatinine ratio (UACR), ocular fundus examination, and diabetes treatment records were collected. An independent sample t-test were used to analyze differences. A Logistic regression analysis was applied to study the independent risk factors of DR. RESULTS: A total of 1282 patients with type 2 DM were enrolled, and 191 cases had DR (14.9%). The age at diabetes onset, education level, alcohol consumption, HbA1c level, UACR level, and hypoglycemic drugs were independent influencing factors for DR. The older the onset of diabetes, the less likely to develop DR (OR: 0.958, 95%CI: 0.942-0.975, P=0.000). Patients were then divided in terms of age at diabetes onset as follows: <50y, 50-59y, 60-69y, and ≥70y. Compared with diabetes onset age <50y, 50-59y (OR: 0.463, 95%CI: 0.306-0.699, P=0.000), 60-69y (OR: 0.329, 95%CI: 0.203-0.535, P=0.000) and ≥70y (OR: 0.232, 95%CI: 0.094-0.577, P=0.002) were at a lower risk of DR. The prevalence of DR was highest in patients with diabetes onset age <50y (29.5%, P<0.05). The HbA1c level (8.67±1.97)% and proportion of insulin injection (52.5%) in patients with diabetes onset <40y were higher than in patients with older diabetes onset age (P<0.05). CONCLUSION: Diabetes onset at an earlier age and uncontrollable high HbA1c level could be independent risk factors for DR.

Study on alkaloid constituents of Stephania tetrandra S. Moore / 药学学报

Fifteen alkaloid compounds were isolated from the 70% aqueous alcohol extract of Stephania tetrandra S. Moore by silica gel, reversed phase silica gel, Sephadex LH-20 column chromatography and semi-preparative high performance liquid chromatography. They were identified as tetrandraside A (1), (Z)-N-formyl-nornuciferin (2), (E)-N-formyl-nornuciferin (3), salutaridine (4), salutaridine N-oxide (5), (E)-3-(4-hydroxy-3-methoxyphenyl)-N-[2-(4-hydroxy-3-methoxyphenyl)ethyl]-2 propenamide (6), dauriporphine (7), sinomenine (8), liriodenine (9), α-magnoflorine (10), (1S)-4'-β-glucosylcoclaurine (11), tetrandrine (12), fangchinoline (13), tetrandrine 2'-β-oxide (14), and tetrandrine 2'-α-oxide (15), respectively, by MS, NMR and single crystal diffraction. Among them, compound 1 is a new alkaloid glycoside. Compounds 2-11 were obtained from this plant for the first time. These compounds showed obvious cytotoxic activity against drug-resistant lung cancer cell line H1299, and compound 9 had the best activity, with an IC50 of 5.38 μmol·L-1.

Psychological flexibility training for career adaptability improvement among second-year middle-school students / 中国学校卫生

Objective@#To test the effectiveness of psychological flexibility training on career adaptability among middle school students who undertook psychological courses based on acceptance commitment therapy and the adolescent mental flexibility model(DNA-V), and to provide a reference plan to improve the mental health of middle school students.@*Methods@#This study recruited 110 junior high school students (60 boys and 50 girls) from a middle-school in Beijing. The students were randomly divided by class into a DNA-V face-to-face course group(offline group n=33), a DNA-V online course group(online group n=40), and a regular school psychology course group(control group n=37). Louise Hayes DNA-V intervention program was condensed into a six-hour middle-school DNA-V psychology curriculum. Using the Avoidance and Fusion Questionnaire for Youth and the Career Adaptability Scale, changes in psychological flexibility and career adaptability were measured before(T1), one week after(T2), and two months after (T3) the intervention.@*Results@#Linear mixed models were used for the analysis, while controlling for demographic variables. Psychological flexibility and career adaptability in the offline group were higher at T2 and T3 than at T1(psychological flexibility t=4.22, 3.11; career adaptablity t=3.05, 4.16, P<0.01), while the difference between T2 and T3 was not statistically significant. The psychological flexibility and career adaptability of the online group were not statistically significant at T1, T2, and T3. The psychological flexibility and career adaptability of the control group increased from T1 to T2(t=4.64, 2.47, P<0.05), but T3 decreased back to a level close to T1.@*Conclusion@#In terms of both psychological flexibility and career adaptability, the DNA-V face-to-face psychology course resulted in a retention period of at least two months.

Role of long noncoding RNA-mediated competing endogenous RNA regulatory network in hepatocellular carcinoma.

Long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) are noncoding RNAs (ncRNAs) that occupy over 90% of the human genome, and their main function is to directly or indirectly regulate messenger RNA (mRNA) expression and participate in the tumorigenesis and progression of malignances. In particular, some lncRNAs can interact with miRNAs as competing endogenous RNAs (ceRNAs) to modulate mRNA expression. Accordingly, these RNA molecules are interrelated and coordinate to form a dynamic lncRNA-mediated ceRNA regulatory network. Mounting evidence has revealed that lncRNAs that act as ceRNAs are closely related to tumorigenesis. To date, numerous studies have established many different regulatory networks in hepatocellular carcinoma (HCC), and perturbations in these ceRNA interactions may result in the initiation and progression of HCC. Herein, we emphasize recent advances concerning the biological function of lncRNAs as ceRNAs in HCC, with the aim of elucidating the molecular mechanism underlying these HCC-related RNA molecules and providing novel insights into the diagnosis and treatment of HCC.

Sex-structured wild and sterile mosquito population models with different release strategies.

In this paper, we propose sex-structured mathematical models in terms of continuous-time differential equations. We investigate the interactive dynamics of the sex-structured wild and sterile mosquitoes from several aspects including the existence of equilibria and their stability. We consider different strategies of releasing the sterile mosquitoes to control mosquitoes in an effective way. In addition, numerical simulations are provided to illustrate the dynamical features of the models.

The spread of influenza-like-illness within the household in Shanghai, China.

High-density urban habitats provide a hotbed for the rapid spread of infectious diseases. School children densely aggregate in classrooms. So schools are high incidence area of infectious diseases. This paper aims at investigating the transmission of influenza-like-illness within households with a school child using a survey study of fourth grade elementary school students in Shanghai, China. We found that the pairwise transmission probability within a household is only 0.172, which implies that the average number of infections caused by a single infectious individual in a household in Shanghai is only 0.304. Thus, the majority of transmission must occur outside of a household for a disease to cause an outbreak.

Effects of Different Stimulators on Proliferation of Peripheral Blood Lymphocyte Subsets / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of different stimultors (PHA, PMA and IL-2) and culture systems (PBMC and whole blood) on the proliferation of human peripheral blood lymphocyte subsets, so as to provide the experimental basis for selecting the appropriate system according to the experimental purposes.@*METHODS@#A total of 10 ml serum samples were collected from healthy volunteers (n=6). The 300 μl whole blood was directly used to detect lymphocyte subsets by flow cytometry. The 400 μl whole blood were inoculated respectively with 3 different stimulators at 37℃ and 5% CO2 for 60 h; Three different stimulators were also added to the PBMC which were isolated from 2 ml whole blood. Then the proliferation ability of lymphocyte subsets was analyzed by flow cytometry.@*RESULTS@#After the PBMC were stimulated with PHA, CD4CD8CD3 lymphocytes were the most subset; The proportion of CD3CD4 T lymphocytes and CD3CD19 B lymphocytes decreased after being stimulated by PMA (P<0.01, P<0.05); the lymphocyte subset ratio had no significant change after being stimulated by IL-2. After the whole blood system was stimulated with PHA, the CD4/CD8 T lymphoblasts were main subsets, the counts of B lymphocytes and NK cells were reduced; after being stimulated with PMA, the number of CD8CD3 T lymphoblast and CD4CD8T lymphocytes increased, the B/NK cells were not distinguished with the surface markers; after the whole blood system was stimulated with IL-2, the proportion of NK cells significantly increased (P<0.05).@*CONCLUSION@#Lymphocyte proliferation stimulated by PMA is the fastest, while the effect of IL-2 on the lymphocyte subset proportion stimulated by IL-2 is the minimal. After being stimulated by PHA the division cycles of lymphocyte are the most.

A new cytotoxic isocoumarin from endophytic fungus Aspergillus versicolor / 中国中药杂志

A new isocoumarin, along with five known ones,were isolated from the fermentation products of an endophytic fungus Aspergillus versicolorby using various chromatographic techniques.Their structures were elucidated on the basis of extensivespectroscopic analysis, including 1D-and 2D-NMR techniques. Compound 1 was evaluated for cytotoxicity against five human tumor cell lines. The results showed that 1 exhibited weak cytotoxicityagainst NB4, SHSY5Y and MCF7 cells with IC₅₀ values of 6.8, 4.3,8.8 μmol•L⁻¹, respectively.

Long-term boron-deficiency-responsive genes revealed by cDNA-AFLP differ between Citrus sinensis roots and leaves.

Seedlings of Citrus sinensis (L.) Osbeck were supplied with boron (B)-deficient (without H3BO3) or -sufficient (10 µM H3BO3) nutrient solution for 15 weeks. We identified 54 (38) and 38 (45) up (down)-regulated cDNA-AFLP bands (transcript-derived fragments, TDFs) from B-deficient leaves and roots, respectively. These TDFs were mainly involved in protein and amino acid metabolism, carbohydrate and energy metabolism, nucleic acid metabolism, cell transport, signal transduction, and stress response and defense. The majority of the differentially expressed TDFs were isolated only from B-deficient roots or leaves, only seven TDFs with the same GenBank ID were isolated from the both. In addition, ATP biosynthesis-related TDFs were induced in B-deficient roots, but unaffected in B-deficient leaves. Most of the differentially expressed TDFs associated with signal transduction and stress defense were down-regulated in roots, but up-regulated in leaves. TDFs related to protein ubiquitination and proteolysis were induced in B-deficient leaves except for one TDF, while only two down-regulated TDFs associated with ubiquitination were detected in B-deficient roots. Thus, many differences existed in long-term B-deficiency-responsive genes between roots and leaves. In conclusion, our findings provided a global picture of the differential responses occurring in B-deficient roots and leaves and revealed new insight into the different adaptive mechanisms of C. sinensis roots and leaves to B-deficiency at the transcriptional level.

Phenylpropanoids and phenylethanol from flowers of Rosa rugosa / 中国中药杂志

A new phenylpropanoid (1), together with seven known ones (2-8), has been isolated from the flowers of Rosa rugosa collected from Shanxi province by using various chromatographic techniques. Compound 1 is a new compound, and it displayed cytotoxicity against NB4, SH-SY5Y, PC3, A549 and MCF7 cell lines with IC₅₀ values of 8.2, 6.2, 4.3, 2.8, and 9.6 µmol · L⁻¹ respectively.

A new flavone from stems of Garcinia bracteata and its anti-TMV activity / 中国中药杂志

A phytochemical investigation on the stems of Garcinia bracteata collected from Xishuangbanna resulted in the isolation of a new flavone. By analysis of the HRESIMS, IR, UV, 1D and 2D NMR spectra, the structure of the new compound was determined as 7-methoxy-4',6-dihydroxy-8-isobutyryl-flavone(1). Compound 1 was also tested for its anti-tobacco mosaic virus(TMV) activity. Results suggested the 1 possessed remarkable anti-TMV activity, with an inhibition rate of 28.2%.

Isoflavones from flower buds of Rosa rugosa and their cytotoxicity / 中草药

Objective: To study the chemical constituents from the flower buds of Rosa rugosa. Methods: The chemical constituents from the flower buds of R. rugosa were isolated by silica gel, MCI-gel resin, Sephadex LH-20 column chromatography and high performance liquid chromatography (HPLC) methods. Their structures were elucidated by spectroscopic methods and physicochemical properties. Results: Three isoflavones, 6,8-dihydroxy-4',7-dimethoxyisoflavone (1), prunetin (2), and pratensein (3) were isolated from the flower buds of R. rugosa. Conclusion: Compound 1 is a new compound named rosa isoflavone. Compounds 2 and 3 are isolated from R. rugosa for the first time. Compound 1 displays the stronger cytotoxicity against A549 and PC3 cells with IC50 values of 2.6 and 3.2 μmol/L, respectively.

Evaluation of the immune response in Shitou geese (Anser anser domesticus) following immunization with GPV-VP1 DNA-based and live attenuated vaccines.

BACKGROUND: Goose parvovirus (GPV) is a highly contagious and deadly disease for goslings and Muscovy ducklings. OBJECTIVES: To compare the differences in immune response of geese immunized with GPV-VP1 DNA-based and live attenuated vaccines. ANIMALS AND METHODS: Shitou geese were immunized once with either 20 µg pcDNA-GPV-VP1 DNA gene vaccine by gene gun bombardment via intramuscular injection, or 300 µg by i.m. injection, or 300 µL live attenuated vaccine by i.m. injection, whereas 300 µg pcDNA3.1 (+) i.m. or 300 µL saline i.m. were used as positive and negative controls, respectively. Each group comprised 28 animals. Peripheral blood samples were collected from 2-210 days after immunization and the proliferation of T lymphocytes, the number of CD4(+) and CD8(+) T cells and the level of IgG assessed. Statistical analysis was performed using a one-way analysis of variance with group multiple comparisons via Tukey's test. RESULTS: The pcDNA-GPV-VP1 DNA and attenuated vaccine induced cellular and humoral responses, and there were no differences between the 20 and 300 µg group in the responses of proliferation of T lymphocyte and the CD8(+) T-cell. However, as to CD4(+) T-cell response and humoral immunity, the 20 µg group performed better than the 300 µg group, which induced better cellular and humoral immunity than live attenuated vaccine. CONCLUSIONS: This study showed that it is possible to induce both cellular and humoral response using DNA-based vaccines and that the pcDNA-GPV-VP1 DNA gene vaccine induced better cellular and humoral immunity than live attenuated vaccine.

Herpes simplex virus 1 counteracts viperin via its virion host shutoff protein UL41.

The interferon (IFN)-inducible viperin protein restricts a broad range of viruses. However, whether viperin plays a role during herpes simplex virus 1 (HSV-1) infection is poorly understood. In the present study, it was shown for the first time that wild-type (WT) HSV-1 infection couldn't induce viperin production, and ectopically expressed viperin inhibited the replication of UL41-null HSV-1 but not WT viruses. The underlying molecular mechanism is that UL41 counteracts viperin's antiviral activity by reducing its mRNA accumulation.

Molecular cloning and characterization of pseudorabies virus EP0 gene.

The pseudorabies virus (PRV) early protein EP0 is a homologue of the herpes simplex virus 1 (HSV-1) immediate-early protein ICP0, which is a multifunctional protein and important for HSV-1 infection. However, the exact function of EP0 is not clear. In this study, using polymerase chain reaction, a 1,104 base-pair sequence of the EP0 gene was amplified from the PRV Becker strain genome and identification of the EP0gene was confirmed by further cloning and sequencing. Bioinformatics analysis indicated that the PRV EP0 gene encoded a putative polypeptide with 367 amino acids. The encoded protein, designated as EP0 contained a conserved RING-finger superfamily domain and was found to be closely related with the herpes virus RING-finger superfamily and was highly conserved among the counterparts encoded by RING-finger genes. Multiple nucleic acid sequence and amino-acid sequence alignments suggested that PRV EP0 showed a relatively higher similarity with EP0-like proteins of genus Varicellovirus than with those of other genera of Alphaherpesvirinae. In addition, phylogenetic analysis showed that PRV EP0 had a close evolutionary relationship with members of genus Varicellovirus, especially bovine herpesvirus 1 (BoHV-1) and BoHV-5. Antigen prediction indicated that several potential B-cell epitopes were located in EP0. Also, subcellular localization analysis demonstrated that EP0 was predominantly localized in the nucleus, suggesting that it might function as a nuclear-targeted protein.