Infectious clone is a useful tool in exploring viral replication and pathogenesis. In order to prevent linear PCV2 cyclization, PCR mutagenesis was used to construct the first molecular clone (pSK-2PCV2) by ligating two copies of the complete PCV2 genome with the pBluescript SK (pSK) vector. In addition, pSK-PCV2 and ds-PCV2 were constructed. PK-15 cells were transfected with above three infectious clones. Indirect immunofluorescence assay (IFA) revealed that the virus antigen mainly localized in infected cell nucleolus and cytoplasm. PCV2 specific nucleotide fragment in cell culture was amplified by RT-PCR. Typical porcine circovirus particles with diameter about 17 nm were also observed by transmission electron microscope (TEM) in the infected cells. The rescued virus sequences from the cultures had 100% homology with the inserting PCV2 genome. The rescued virus shared similar properties with that of the parental virus. The study establishes a platform for further research on the virus molecular biology and pathogenicity.
Animals ; Cell Line ; Circoviridae Infections ; virology ; Circovirus ; genetics ; growth & development ; pathogenicity ; physiology ; Cloning, Molecular ; DNA, Viral ; genetics ; physiology ; Recombination, Genetic ; genetics ; Swine ; Transfection ; Virulence ; Virus Replication

Objective:To discuss the effect of lumbar disc herniation of motility kinesiophobia during convalescence which intervened by Chinese medicine nursing.Methods:A total of 172 patients with lumbar disc herniation treated at TCM Rehabitation Department at Three Gorges Hospital Affiliated to Chongqing University from May 1, 2017 to January 30, 2019 were selected and divided into control group and intervention group by random digits table method with 86 cases each. The control group was given routine psychological nursing, and the intervention group was given emotional nursing of traditional Chinese medicine combined with cognitive behavior intervention. The scores of anxiety, depression and kinesiophobia, the quality of life, the completion and accuracy of rehabilitation training before and after intervention were compared between the two groups.Results:There was no significant difference in the scores of anxiety, depression and kinesiophobia, the quality of life, the completion and accuracy of rehabilitation training before intervention between the two groups( P>0.05). The scores of anxiety, depression at 1,2,3 weeks after intervention was (54.61±7.23), (49.98±6.55), (45.03±6.21) and (55.16±7.53), (50.96±6.45), (46.31±5.72) in the intervention group, (59.69±7.52), (56.92±6.79), (53.62±7.28) and (60.65±7.96), (56.87±6.80), (52.80±5.92) in the control group, the differences were significant( Fgroup values were 10.157, 9.138, P<0.05). The scores of kinesiophobia, the quality of life at 1,2,3 weeks after intervention was (39.66±4.02), (30.58±4.02), (24.38±3.87) and (43.83±2.74), (56.72±3.81), (75.27±5.83)in the intervention group, (43.71±6.87), (37.83±6.81), (33.95±2.86) and (42.50±3.92), (46.05±3.95), (51.62±4.64) in the control group, the differences were significant( Fgroup values were 30.718, 30.938, P<0.01). The completion and accuracy of rehabilitation training at 1, 2, 3 weeks after intervention was (69.08±1.83), (77.05±2.04), (87.34±3.16) and (60.32±2.19), (70.42±3.22), (83.93±2.74) in the intervention group, (54.23±4.22), (62.72±2.65), (73.51±5.41) and (48.24±5.07), (59.77±4.13), (75.64±3.33) in the control group, the differences were significant( Fgroup values were 28.471, 25.139, P<0.01). Conclusions:Emotion nursing of traditional Chinese medicine combined with cognitive behavioral intervention is helpful to relieve the anxiety and depression of patients with lumbar disc herniation, effectively reduce the kinesiophobia of patients during rehabilitation, improve the quality of life of patients, improve the completion and accuracy of rehabilitation training of patients, and contribute to the improvement of rehabilitation effect and the realization of good treatment outcome.

Objective:To explore the mechanism of social support and self-efficacy in rehabilitation exercise in the relationship between psychological resilience and kinesiophobia in hip fracture patients.Methods:Totally 284 hip fracture patients treated at the Department of Trauma and Orthopedics, Binzhou Medical University Hospital from May 2022 to April 2023 were selected by convenience sampling. Data were collected using the general information questionnaire, the Connor-Davidson Resilience Scale (CD-RISC), the Social Support Rating Scale (SSRS), the Self-Efficacy Rehabilitation Outcome Scale (SER), and the Tampa Scale for Kinesiophobia (TSK). Process 4.1 was used for chain mediation effect analysis.Results:The scores for the 284 hip fracture patients were as follows: CD-RISC (58.72±7.77), SSRS (29.70±5.11), SER (91.23±8.41), and TSK (38.07±3.65). Psychological resilience, social support, self-efficacy in rehabilitation exercise, and kinesiophobia were all significantly correlated pairwise (all P<0.01). Social support and self-efficacy in rehabilitation exercise had a chain mediation effect between psychological resilience and kinesiophobia, with mediation effect values of -0.020 and -0.079, accounting for 17.52% and 67.70% of the total effect, respectively. Conclusions:Psychological resilience directly influences the level of kinesiophobia in hip fracture patients and has an indirect effect through the chain mediation of social support and self-efficacy in rehabilitation exercise.

Hepatocellular carcinoma(HCC)is one of the most common tumor types and remains a major clinical challenge.Increasing evidence has revealed that mitophagy inhibitors can enhance the effect of chemotherapy on HCC.However,few mitophagy inhibitors have been approved for clinical use in humans.Pyrimethamine(Pyr)is used to treat infections caused by protozoan parasites.Recent studies have reported that Pyr may be beneficial in the treatment of various tumors.However,its mechanism of action is still not clearly defined.Here,we found that blocking mitophagy sensitized cells to Pyr-induced apoptosis.Mechanistically,Pyr potently induced the accumulation of autophagosomes by inhibiting autophagosome-lysosome fusion in human HCC cells.In vitro and in vivo studies revealed that Pyr blocked autophagosome-lysosome fusion by upregulating BNIP3 to inhibit synaptosomal-associated protein 29(SNAP29)-vesicle-associated membrane protein 8(VAMP8)interaction.Moreover,Pyr acted synergistically with sorafenib(Sora)to induce apoptosis and inhibit HCC proliferation in vitro and in vivo.Pyr enhances the sensitivity of HCC cells to Sora,a common chemotherapeutic,by inhibiting mitophagy.Thus,these results provide new insights into the mechanism of action of Pyr and imply that Pyr could potentially be further developed as a novel mitophagy inhibitor.Notably,Pyr and Sora combination therapy could be a promising treatment for malignant HCC.