Extracellular vesicles (EVs) are phospholipid bilayer vesicles actively secreted by cells, that contain a variety of functional nucleic acids, proteins, and lipids, and are important mediums of intercellular communication. Based on their natural properties, EVs can not only retain the pharmacological effects of their source cells but also serve as natural delivery carriers. Among them, plant-derived nanovesicles (PNVs) are characterized as natural disease therapeutics with many advantages such as simplicity, safety, eco-friendliness, low cost, and low toxicity due to their abundant resources, large yield, and low risk of immunogenicity in vivo. This review systematically introduces the biogenesis, isolation methods, physical characterization, and components of PNVs, and describes their administration and cellular uptake as therapeutic agents. We highlight the therapeutic potential of PNVs as therapeutic agents and drug delivery carriers, including anti-inflammatory, anticancer, wound healing, regeneration, and antiaging properties as well as their potential use in the treatment of liver disease and COVID-19. Finally, the toxicity and immunogenicity, the current clinical application, and the possible challenges in the future development of PNVs were analyzed. We expect the functions of PNVs to be further explored to promote clinical translation, thereby facilitating the development of a new framework for the treatment of human diseases.

Objective Colon-targeting capsules based on gastric pellets and enteric pellets were prepared from Baizhu Huanglian prescription. The formulation composition and preparation process were optimized and the in-vitro release characteristics were investigated. Methods Optimum formulation composition and process parameters of Baizhu Huanglian pellets were screened out by single factor experiment and orthogonal design. The pellets core were prepared by extrusion-spheronization technique and coated in the fluid bed using bottom spray coating technique. To investigate the effect of coating level of the isolation layer, the proportion of polymer, the amount of plasticizer and weight gain of enteric coating on the release behavior of the enteric pellets. The pellets release behavior was fitted by model as well. Results The prescription of gastric pellets was drug loading 50%, PVPP 5%, MCC to lactose 1∶2 and wetting agent 40%. The process parameters were extrusion frequency 20 Hz, rounding speed 500 r/min and rounding time 5 min. The prescription of enteric pellets was drug loading 27%, PVPP 5%, MCC to lactose 5∶2, wetting agent 30% and adhesive 20%. The process parameters were extrusion frequency 20 Hz, rounding speed 700 r/min and rounding time 7 min. For enteric coating layer, the coating mixture of EUDRAGIT®L30D-55 to EUDRAGIT® FS30D was 1∶2. The amount of plasticizer was 10%. The increased weight of coating layer was 15%. The release time of enteric pellets in-vitro was up to 24 hours. The release behavior of the pellets conforms to the Higuchi model. Conclusion The colon targeting capsule of Baizhu Huanglian pellets were successfully prepared and showed the characteristics of sustained release and colon targeting.

Objective:To explore the causes and precaution procedures of olfactory dysfunction after transsphenoidal surgery for pituitary lesions.Methods:The clinical data of 72 patients underwent transsphenoidal surgery for pituitary lesions in our hospital from November 2016 to May 2018 were summarized. Resection under microscope was performed in 27 patients and resection under endoscope in 45 patients. The olfactory function of patients with postoperative olfactory disorders was evaluated one and 6 months after surgery. Bilateral nasal passages were examined by nasal endoscopy 3-6 months after surgery. Whether intraoperative operations were related to olfactory impairment was analyzed.Results:There were seven hyposmic/anosmic patients; three patients (6.7%) were operated via endoscopic approach and four patients (14.8%) were operated via microscopic transsphenoidal approach; incidence of olfactory dysfunction between the two groups showed no statistical difference ( P>0.05). One month after operation, two patients had a slight decrease in olfactory function, 4 had a severe decrease in olfactory function and one had anosmia. Six months after the operation, two patients with severe hypoxia got improvment, but failed to return to normal level; and the rest of the patients showed no significant improvement. Three-6 months after operation, nasal endoscopic examination found that all patients had healed nasal wound, two patients had large volume of nasal mucus secretion, and 3 patients had postoperative nasal mucosa adhesion. In these 72 patients,cauterization of the mucosa around the sphenoid ostium was performed in 6 patients, placement of the speculum above the sphenoid ostium was performed in 2 patients, and olfactory damage was noted in all the 8 patients. Conclusions:Olfactory dysfunction after transsphenoidal surgery for pituitary lesions is common, and attention should be paid.Placement of the speculum above the sphenoid ostium and cauterization of the mucosa around the sphenoid ostium might be the main reasons for olfactory damage.

Porcine deltacoronavirus (PDCoV) is a newly emerging enteropathogenic swine coronavirus causing acute diarrhea and vomiting in pigs. The apoptosis of ST cells induced by PDCoV infection was studied in this research. In ST cells, caspase activity assay showed that the activity of caspase 3, caspase 8 and caspase 9 increased significantly with the infection of PDCoV, but not observed in UV irradiated PDCoV-infected cells, indicating that PDCoV infection activated both endogenous and exogenous apoptotic pathways in ST cells, and the induction of apoptosis depended on viral replication. To further investigate the endogenous apoptosis induced by PDCoV, cytochrome C and apoptosis-inducing factors in cytoplasm and mitochondria were detected. Compared with normal cells, the amount of cytochrome C released from mitochondria to cytoplasm increased significantly in PDCoV-infected cells, and the release increased with the prolongation of infection, while the apoptosis-inducing factor was always localized to mitochondria, suggesting that PDCoV induced apoptosis was initiated through caspase-dependent mitochondrial apoptosis pathway by promoting cytochrome C in the mitochondrial membrane gap into the cytosol. In conclusion, this study reveals the mechanism of PDCoV inducing apoptosis.
Animals ; Apoptosis ; Coronavirus ; Coronavirus Infections ; Mitochondria ; Swine ; Swine Diseases

A novel protein encoded by the open reading frame 4 (ORF4) was recently discovered in porcine circovirus type 2 (PCV2). However, little is known about the interaction proteins of ORF4 which hindered better understanding the biological functions of ORF4 in the life cycle of PCV2. In the present study, the ORF4 was inserted into the multiple cloning site of pCMV-N-Flag-GST, yielding recombinant plasmid pCMV-N-Flag-GST-ORF4. The recombinant plasmid was transfected into 293T cells and the intracellular interaction complex of ORF4 were enriched and separated by GST pull-down and SDS-PAGE, sequentially. The potential interacting proteins of PCV2 ORF4 were stained with silver and identified by mass spectrometry (MS). Finally, five candidate ORF4-interacting proteins, including Serine/threonine-protein phosphatase 6 catalytic subunit, alpha cardiac muscle 1, actin, SEC14-like protein 5 and myosin 9 were identified. These results would benefit a better understanding of the biological function of ORF4 in PCV2 infected cells.
Animals ; Circoviridae Infections ; Circovirus ; HEK293 Cells ; Humans ; Mass Spectrometry ; Open Reading Frames ; Swine ; Viral Proteins

Several putative transcription factor binding sites (TFBSs) exist in the PCV2 rep gene promoter. To explore if porcine circovirus type 2 (PCV2) could regulate the viral replication by using these TFBSs, we conducted electrophoretic mobility shift assay (EMSA), DNA-pull down and liquid chromatography-tandem mass spectrometric (LC-MS/MS) assays. EMSA confirmed the binding activity of the rep gene promoter with nuclear proteins of host cells. DNA-pull down and LC-MS/MS identified the porcine transcription factor AP-2δ (poTFAP2δ) could bind the PCV2 rep gene promoter. Dual-luciferase reporter assay, quantitative real-time PCR, Western blotting and indirect immunofluorescent assay demonstrated that poTFAP2δ could not only promote the activity of the rep gene promoter, but also enhance the transcription/translation activity of the rep/cap gene and the virus titer of PCV2 during the entire life cycle of PCV2 infection. This study revealed the molecular mechanism of PCV2 using host proteins to enhance the viral replication, provided a new perspective for studying the pathogenic mechanism of PCV2 from virus and host interactions, and provided a theoretical basis for developing highly effective PCV2 vaccines.
Animals ; Cell Line ; Chromatography, Liquid ; Circoviridae Infections ; Circovirus ; DNA Helicases ; Diabetes Mellitus, Type 2 ; Promoter Regions, Genetic ; Swine ; Tandem Mass Spectrometry ; Transcription Factor AP-2 ; Virus Replication

Porcine circovirus type 2 (PCV2) can cause immunosuppression on herds. PCV2, as an essential pathogen of PCV2-systemic disease (PCV2-SD), has caused considerable economic losses in pig industry worldwide. Here we review and address the evolution, viral protein and immunolesion of PCV2 and preventive techniques of PCV2-SD.
Animals ; Circoviridae Infections ; veterinary ; Circovirus ; genetics ; Phylogeny ; Swine ; Swine Diseases ; virology ; Viral Proteins ; genetics

ObjectiveTo investigate the clinical effects of applying low abdominal superficial artery flap to repair skin and soft tissues defects in upper limbs.MethodsTotal 42 cases with soft tissues defects were recruited in this study.Seven cases were repaired using skin flaps with circumfiexa ilium superficial artery.Five cases were repaired using osteocutaneous flaps based on circumfiexa ilium superficial artery.Eleven cases were repaired using flaps with superficial epigastric artery.Seven cases were repaired using flaps with circumfiexa ilium superficial artery,lateral branches of superficial epigastric artery and musculocutaneous perforators of the fourth lumbar artery.Six cases were repaired using flaps with origin of circumfiexa ilium superficial artery,medial and lateral branch of superficial epigastric artery,and the deep and cutaneous branch of circumfiexa ilium superficial artery.ResultsAfter surgery,forty-two flaps all survived.Flaps areas were between 5 cm × 6 cm-9 cm × 30 cm.Followed up 6-24 months,the appearance,texture,color and function of flaps were recovered well and did not need to trim again.ConclusionLow abnominal superficial artery flap could be used to repair skin and soft tissues,also its texture is so thin and areas can be direct sutures,which,is an ideal method to forearm repairment.

Using the CT data, we have constructed the finite element models of human femur distally amputated at high-position, middle-position and low-position, along with distally osseointegrated implant under the maximal load during a normal walking cycle. Results of finite element analysis revealed: the maximal stress of implant is produced near the exit of the amputated limb, where the fatigue breakpoint caused by cyclic stress would take place. With the ascending of truncated position, the peri-implant interfacial stress of bone increases. There is severe stress-shielding at the bone-implant interface, and there is concentration of stress at the end of implant and at the 3/4 point of femur, which would lead to bone loss and bone resorption and would shorten the longevity of implant. The results also showed that the curvature of natural bone has notable effect on the stress distribution, which should not be neglected. These data may provide reliable reference for the design and research of osseointegarted artificial limb.
Artificial Limbs ; Computer Simulation ; Femur ; physiology ; Finite Element Analysis ; Humans ; Imaging, Three-Dimensional ; Models, Biological ; Osseointegration ; physiology ; Prosthesis Design ; Stress, Mechanical

Objective To explore the diagnostic value of telomerase activity (TA) and hTERT expression in pleural effusions. Methods According to the final diagnosis, 51 patients with pleural effusion were classified into two groups: malignant group (n=27) and benign group (n=24). Telomerase activity in pleural effusion was measured by TRAP-ELISA. Then the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of telomerase activity for diagnosing pleural effusion were calculated. hTERT in pleural effusion was detected by western blotting analysis. Results TA assay showed that 24 of 27 cases of malignant pleural effusion were found to possess telomerase activity, while 1 of 24 cases of benign one had telomerase activity. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of TA assay for diagnosing pleural effusion were 88.9%, 95.8%, 96%, 88.5%, and 92.2%, respectively. hTERT test showed that 8 of 27 cases of malignant pleural effusion had hTERT expression, while none of 12 cases of benign one had hTERT expression. There was significant difference in diagnosing pleural effusion between two methods (? 2=19.64,P=9.4?10 -5 ). Conclusion Tolomerase activity assay may act as a routine diagnostic method for malignant pleural effusions, while hTERT test may serve as complementary one.