Objective To investigate the effect of acidic serine protease ASPNJ on the expression of heat shock protein HSP90, 60 and 27 in human chronic myeloid leukemia K562 cells, in order to reveal the related mechanism of anti leukemic effects of ASPNJ. Methods K562 leukemia cell lines were cultured in vitro and treated with ASPNJ alone or in combination with chemotherapeutic agents. Western blot and RT-PCR were used to detect the changes of HSP90, 60 and 27 gene expressions in levels of total protein and membrane protein, as well as in mRNA levels. Results ASPNJ showed different effects on the expression of HSPs in total protein and membrane protein levels and had some modified effect on HSPs in total protein or membrane protein levels. Effects of ASPNJon expression of HSPs mRNA were not apparent, but HSPs mRNA were apparently lower in the ASPNJ and doxorubicin combination group than that in the ASPNJ alone or doxorubicin alone groups. Conclusion The mechanism of ASPNJ on the inhibitory effect of leukemia cells proliferation and the promoting effects on chemotherapeutic drugs may involve some complicated correlations with the effect of ASPNJ on the expression of HSPs and the modification of HSPs proteins.

Proteins, which exist mainly in linear form in vivo, are easily affected by the change of ambient temperature and pH. The application of proteins (enzymes) in the fields of industrial catalyzing, food manufacturing and medicine are restricted due to their properties. The cyclic structure of natural cyclic peptides confers high thermal stability on itself; such mechanism can be referred to in further enhancement of the thermal stability and transformation of the structure of enzymes. This article reviewed the latest progress in the domestic and international studies on protein cyclization and summarized the traditional methods (such as protein trans-splicing, expressed protein ligation and sortase-catalyzed transpeptidation) in protein cyclization. A novel method based on SpyTag/SpyCather-mediated enzyme cyclization was discussed in more detail.
Cyclization ; Peptides, Cyclic ; chemistry ; Protein Processing, Post-Translational ; Proteins ; chemistry

Objective To investigate the influence of perceptions and emotional attitudes on the public's willingness to organ donation and its path of promotion. Methods The mediation effect and structural equation models were established through the convenience sampling method and with ABC attitude model as the theoretical basis to analyze the influence of perceptions and emotional attitudes on the public's willingness to organ donation and the path of promotion. Results Among 4 565 investigated subjects, 621 subjects expressly stated that they were not willing to donate their organs after the death, 701 subjects were willing to donate their organs after the death, but only 259 investigated subjects signed the informed content card of organ donation. The differences in the subjects' willingness to donate their organs were statistically significant in terms of different genders, ages, religious beliefs, places of residence and educational degrees (all P < 0.05). The overall Cronbach's α coefficient of the questionnaire was 0.781, KMO=0.842, with good reliability and validity. In the structural equation model, the path coefficient of perceptions on the willingness to donation was 0.39, while that of attitudes on the willingness was 0.25. As such, perceptions and emotional attitudes had positive impacts on the willingness to donate the organs. The results of the mediation effect model indicated that attitudes played significant mediation effects in the causality relationship of perceptions on the willingness to donate organs, and the mediation effect value was 0.035(P < 0.01). The awareness degree of organ donation was the largest determinant to the perception factor, and the path coefficient on the willingness to donation was 0.20. The sense of social honor was the largest determinant to the attitude factor, and the path coefficient was 0.16. Conclusions Both perceptions and emotional attitudes positively impact the willingness to donate organs. The awareness degree of organ donation is the largest determinant to the perception factor, while the sense of social honor is the largest determinant to the attitude factor. To improve the public's perception level towards the organ donation and increase the public's sense of social honor towards organ donation contributes to the improvement of the public's willingness to donate organs.

Objective To explore the important factors influencing organ donation willingness and coordination effect of organ donation coordinators. Methods A questionnaire survey was conducted among 349 national organ donation coordinators by convenience sampling, including 145 males and 204 females, aged 27 (23, 36) years. Multiple linear regression and disordered logistic regression were used to investigate the important factors influencing the willingness to donate organs and coordination effects. Results Among 349 organ donation coordinators, 146 (41.8%) were willing to donate organs, including 101 (28.9%) who had signed the consent card for organ donation. Adequate awareness of organ donation laws, high education level, marital experience, and good self-perceived health status all showed positive effects on organ donation willingness of organ donation coordinators (all P < 0.05). High income, long length of service as organ donation coordinators, full-time mode of employment, high willingness to donate organs, and adequate awareness of donation conditions and donation procedures all showed positive effects on the coordination effect of organ donation coordinators (all P < 0.05). Conclusions The willingness to donate organs is increased as the higher awareness of organ donation laws of organ donation coordinators, while enhancing the willingness to donate organs of organ donation coordinators exerts positive impact upon improving the coordination effect of organ donation coordination. Therefore, an all-round organ donation coordinator training system should be established to improve the success rate of organ donation advocacy and promote the development of organ donation.

Objective To investigate the effects of fentanyl citrate on pain in aged rats with hip fracture and its underlying mechanisms.Methods Thirty aged rats with hip fracture were divided in-to model group(n=10),fentanyl citrate group(n=10),and fentanyl citrate+Sar-SP group(n=10).Additionally,healthy rats from the same period were included as normal group(n=10).Rats in the fentanyl citrate group received intravenous tail injection of 10 pg/kg fentanyl citrate,while those in the fentanyl citrate+Sar-SP group underwent intrathecal injection of 10 μL Sar-SP for fenta-nyl citrate intervention.Rats in the normal and model groups received intravenous tail injection of an equal volume of saline.Pain threshold,weight-bearing capacity,temperature,and dorsoventral thick-ness of the hind paws were measured before treatment(0h)and at 1,6,24,and 168 h post-treat-ment.Enzyme-linked immunosorbent assay(ELISA)was used to detect levels of tumor necrosis fac-tor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β),and glutamic acid(Glu)in the spinal cord tissue of rats.Immunohistochemical staining was employed to observe the positive expression rates of ionized calcium-binding adapter molecule 1(Iba-1),glial fibrillary acidic protein(GFAP),and substance P(SP)in the spinal cord tissue.Real-time fluorescent quantitative polymerase chain re-action(qRT-PCR)was conducted to detect the relative expression levels of SP and tachykinin re-ceptor 1(TACR1)mRNA in the hippocampus tissue of rats.Western blot analysis was performed to assess the relative expression levels of vimentin(VIM),nuclear receptor corepressor 1(NOCR),ciliary neurotrophic factor receptor(CNTFR),and epidermal growth factor receptor(EGFR)pro-teins in the spinal cord tissue.Results Compared with the normal group,rats in the model group exhibited decreased pain threshold and weight-bearing capacity,increased hind paw temperature and dorsoventral thickness,as well as elevated levels of TNF-α,IL-6,IL-1 β,and Glu,increased posi-tive expression rates of Iba-1 and GFAP,heightened positive expression rate and relative mRNA ex-pression levels of SP and TACR1,and augmented relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 0 h before treatment and at 1,6,24,and 168 h post-treatment(P＜0.05).Compared with the model group,rats in the fentanyl citrate group showed increased pain threshold and weight-bearing capacity,decreased hind paw temperature and dorsoventral thickness,as well as reduced levels of TNF-α,IL-6,IL-1β,and Glu,decreased posi-tive expression rates of Iba-1 and GFAP,lowered positive expression rate and relative mRNA expres-sion levels of SP and TACR1,and decreased relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 1,6,24,and 168 h post-treatment(P＜0.05).Compared with the fentanyl citrate group,rats in the fentanyl citrate+Sar-SP group demonstrated decreased pain threshold and weight-bearing capacity,increased hind paw temperature and dorso-ventral thickness,elevated levels of TNF-α,IL-6,IL-1 β,and Glu,increased positive expression rates of Iba-1 and GFAP,heightened positive expression rate and relative mRNA expression levels of SP and TACR1,and elevated relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 1,6,24,and 168 h post-treatment(P＜0.05).Conclusion The im-provement in pain in aged rats with hip fracture by fentanyl citrate may be associated with the reduc-tion of SP expression in the spinal cord that inhibits glial cell activation and exhibits anti-inflammato-ry effects.

Protein arginine methylation is an important post-translational modification(PTM)mediated by protein arginine methyltransferase(PRMTs),which is closely related to the occurrence and development of many diseases.Methylation of arginine is closely related to inflammatory diseases and fracture healing.Decreased expression of PRMTs can lead to delayed or even non-healing of frac-tures.Both PRMT5 and PRMT6 play an important role in fracture healing and are closely related to the PI3K-AKT and NF-κB pathways.Exploration the relationship between protein arginine methyla-tion and fracture healing can provide a new way to prevent delayed or even non-healing fracture.

Objective To investigate the effects of fentanyl citrate on pain in aged rats with hip fracture and its underlying mechanisms.Methods Thirty aged rats with hip fracture were divided in-to model group(n=10),fentanyl citrate group(n=10),and fentanyl citrate+Sar-SP group(n=10).Additionally,healthy rats from the same period were included as normal group(n=10).Rats in the fentanyl citrate group received intravenous tail injection of 10 pg/kg fentanyl citrate,while those in the fentanyl citrate+Sar-SP group underwent intrathecal injection of 10 μL Sar-SP for fenta-nyl citrate intervention.Rats in the normal and model groups received intravenous tail injection of an equal volume of saline.Pain threshold,weight-bearing capacity,temperature,and dorsoventral thick-ness of the hind paws were measured before treatment(0h)and at 1,6,24,and 168 h post-treat-ment.Enzyme-linked immunosorbent assay(ELISA)was used to detect levels of tumor necrosis fac-tor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β),and glutamic acid(Glu)in the spinal cord tissue of rats.Immunohistochemical staining was employed to observe the positive expression rates of ionized calcium-binding adapter molecule 1(Iba-1),glial fibrillary acidic protein(GFAP),and substance P(SP)in the spinal cord tissue.Real-time fluorescent quantitative polymerase chain re-action(qRT-PCR)was conducted to detect the relative expression levels of SP and tachykinin re-ceptor 1(TACR1)mRNA in the hippocampus tissue of rats.Western blot analysis was performed to assess the relative expression levels of vimentin(VIM),nuclear receptor corepressor 1(NOCR),ciliary neurotrophic factor receptor(CNTFR),and epidermal growth factor receptor(EGFR)pro-teins in the spinal cord tissue.Results Compared with the normal group,rats in the model group exhibited decreased pain threshold and weight-bearing capacity,increased hind paw temperature and dorsoventral thickness,as well as elevated levels of TNF-α,IL-6,IL-1 β,and Glu,increased posi-tive expression rates of Iba-1 and GFAP,heightened positive expression rate and relative mRNA ex-pression levels of SP and TACR1,and augmented relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 0 h before treatment and at 1,6,24,and 168 h post-treatment(P＜0.05).Compared with the model group,rats in the fentanyl citrate group showed increased pain threshold and weight-bearing capacity,decreased hind paw temperature and dorsoventral thickness,as well as reduced levels of TNF-α,IL-6,IL-1β,and Glu,decreased posi-tive expression rates of Iba-1 and GFAP,lowered positive expression rate and relative mRNA expres-sion levels of SP and TACR1,and decreased relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 1,6,24,and 168 h post-treatment(P＜0.05).Compared with the fentanyl citrate group,rats in the fentanyl citrate+Sar-SP group demonstrated decreased pain threshold and weight-bearing capacity,increased hind paw temperature and dorso-ventral thickness,elevated levels of TNF-α,IL-6,IL-1 β,and Glu,increased positive expression rates of Iba-1 and GFAP,heightened positive expression rate and relative mRNA expression levels of SP and TACR1,and elevated relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 1,6,24,and 168 h post-treatment(P＜0.05).Conclusion The im-provement in pain in aged rats with hip fracture by fentanyl citrate may be associated with the reduc-tion of SP expression in the spinal cord that inhibits glial cell activation and exhibits anti-inflammato-ry effects.

Protein arginine methylation is an important post-translational modification(PTM)mediated by protein arginine methyltransferase(PRMTs),which is closely related to the occurrence and development of many diseases.Methylation of arginine is closely related to inflammatory diseases and fracture healing.Decreased expression of PRMTs can lead to delayed or even non-healing of frac-tures.Both PRMT5 and PRMT6 play an important role in fracture healing and are closely related to the PI3K-AKT and NF-κB pathways.Exploration the relationship between protein arginine methyla-tion and fracture healing can provide a new way to prevent delayed or even non-healing fracture.

BACKGROUND: Nowadays, biological tissue engineering is a growing field of research. Biocompatibility is a key indicator for measuring tissue engineering biomaterials, which is of great significance for the replacement and repair of damaged tissues. METHODS: In this study, using gelatin, carboxymethyl chitosan, and sodium alginate, a tissue engineering material scaffold that can carry cells was successfully prepared. The material was characterized by Fourier transforms infrared spectroscopy. In addition, the prepared scaffolds have physicochemical properties, such as swelling ratio, biodegradability.we observed the biocompatibility of the hydrogel to different adult stem cells (BMSCs and ADSCs) in vivo and in vitro. Adult stem cells were planted on gelatin-carboxymethyl chitosan-sodium alginate (Gel/SA/CMCS) hydrogels for 7 days in vitro, and the survival of stem cells in vitro was observed by live/died staining. Gel/SA/CMCS hydrogels loaded with stem cells were subcutaneously transplanted into nude mice for 14 days of in vivo culture observation. The survival of adult stem cells was observed by staining for stem cell surface markers (CD29, CD90) and Ki67. RESULTS: The scaffolds had a microporous structure with an appropriate pore size (about 80 lm). Live/died staining showed that adult stem cells could stably survive in Gel/SA/CMCS hydrogels for at least 7 days. After 14 days of culture in nude mice, Ki67 staining showed that the stem cells supported by Gel/SA/CMCS hydrogel still had high proliferation activity. CONCLUSION Gel/SA/CMCSs hydrogel has a stable interpenetrating porous structure, suitable swelling performance and degradation rate, can promote and support the survival of adult stem cells in vivo and in vitro, and has good biocompatibility. Therefore, Gel/SA/CMCS hydrogel is a strong candidate for biological tissue engineering materials.

Objective:To investigate the changes of hippocampal gray matter volume and expression of candidate immune related genes in a rat model of schizophrenia established by repeated administration of dizocilpine(MK-801).Methods:Thirty SPF grade Sprague-Dawley male rats at postnatal day 28 were randomly divided into MK-801 medium-dose (0.25 mg/kg) group, MK-801 high-dose(0.50 mg/kg) group and normal saline (5 mL/kg) group according to random number table method, with 10 in each group.Rats were given continuous intraperitoneal administration according to grouping once a day for 14 days.Open field test, novel object recognition test and Y-maze test were used at postnatal day 60 to detect spontaneous activity, exploration ability, anxiety level, object recognition memory ability and spatial working memory of rats, respectively.At postnatal day 67, structural magnetic resonance imaging was used to detect the changes of hippocampal gray matter volume in rat.And at postnatal day 70, qRT-PCR was used to detect the expression of candidate immune-related genes in rat hippocampus.SPSS 25.0 was used for statistical analysis, one-way ANOVA was used for comparison among multiple groups, and Tukey test was used for further pairwise comparisons.Results:(1)The behavioral results showed that there were significant differences in the total movement distance, central area activity time, novel object recognition index, and spontaneous correct alternation rate among the three groups ( F=11.15, 10.11, 13.62, 11.99, all P<0.05). The total movement distances in MK-801 medium-dose group and MK-801 high-dose group ((21.44±2.17) m, (22.87±1.96)m) were higher than that in the normal saline group ((18.70±1.88) m) (both P<0.05). The activity time of the central area in the MK-801 medium-dose group and MK-801 high-dose group((3.24±1.58) s, (2.50±1.32) s) were lower than that of the normal saline group ((6.05±2.48)s) (both P<0.01). Novel object recognition indexes in the MK-801 medium-dose group and MK-801 high-dose group((56.10±3.99)%, (54.00±6.41)%) were both lower than that in the normal saline group ((65.90±5.65)%)(both P<0.01), and the rates of spontaneous correct alternation ((54.60±7.03)%, (51.60±8.84)%) in the two groups were lower than that of the normal saline group ((68.40±8.57)%) (both P<0.01). (2) The results of structural magnetic resonance imaging showed that there were significant differences in the volume of hippocampal gray matter among the three groups ( F=9.24, P<0.001). The volumes of hippocampal gray matter in MK-801 medium-dose group and MK-801 high-dose group were lower than that in normal saline group(both P<0.001). (3)By constructing protein-protein interaction network, four candidate immune related genes were screened out: neuropeptide Y (NPY), somatostatin (SST), cholecystokinin (CCK) and tachykinin 1 (TAC1). The results showed that the mRNA expression levels of NPY, SST and CCK in the hippocampus of the three groups were significantly different ( F=11.41, 10.43, 5.85, all P<0.05), but there was no statistical difference in the TAC1 mRNA expression level ( F=0.08, P>0.05). The mRNA levels of NPY, SST and CCK in the hippocampus of rats in the MK-801 high-dose group were lower than those in the normal saline group (all P<0.05). Conclusion:Both medium dose and high dose MK-801 administration can reduce the volume of hippocampal gray matter in schizophrenia model rats, but they have different effects on the expression of hippocampal immune related genes, of which high dose administration has a greater effect.