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.

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 into 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 μg/kg fentanyl citrate, while those in the fentanyl citrate + Sar-SP group underwent intrathecal injection of 10 μL Sar-SP for fentanyl 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 thickness of the hind paws were measured before treatment (0 h) and at 1, 6, 24, and 168 h post-treatment. Enzyme-linked immunosorbent assay (ELISA) was used to detect levels of tumor necrosis factor-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 reaction (qRT-PCR) was conducted to detect the relative expression levels of SP and tachykinin receptor 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) proteins 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-1β, 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 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 positive expression rates of Iba-1 and GFAP, lowered positive expression rate and relative mRNA expression 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 dorsoventral thickness, elevated levels of TNF-α, IL-6, IL-1β, 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 improvement in pain in aged rats with hip fracture by fentanyl citrate may be associated with the reduction of SP expression in the spinal cord that inhibits glial cell activation and exhibits anti-inflammatory 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 fractures. 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 methylation and fracture healing can provide a new way to prevent delayed or even non-healing fracture.

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.

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 study the impact of donor left hepatic vein classification and the reconstruction methods on hepatic venous outflow obstruction (HVOO) after pediatric living-donor liver transplantation using left lateral liver segments.Methods:A retrospective study was performed on the clinical data of 653 children recipients who underwent living-donor liver transplantation with left lateral liver segments from January 2014 to December 2020 at Tianjin First Central Hospital. There were 309 males and 344 females, aged 7.0 (6.0, 10.0) months, with an age range of 3-121 months. Based on the left hepatic vein on preoperative donor enhancement CT as well as the intraoperative reconstruction methods, the recipients were divided into 3 groups: type Ⅰ group ( n=514), anastomosis using a single opening was performed directly between the donor and the recipient; type Ⅱ group ( n=118), angioplasty was performed on two adjacent recipient venous orifices before anastomosis, and type Ⅲ group ( n=21), an interposition vessel was anastomosed to two widely spaced openings or the two veins were anastomosed separately. The preoperative general status of the patient, postoperative HVOO incidences, and graft and recipient survival rates were compared among the three groups. The patients were followed up by outpatient reexamination or telephone. Results:Graft to recipient weight ratio in the type Ⅲ group was smaller than that in the type Ⅰ group and the type Ⅱ group ( P<0.05). For all the 653 patients, the incidence of postoperative HVOO was 4.59% (30/653), with the incidences of HVOO in the 3 groups of patients were 4.1% for the type Ⅰ group (21/514), 5.1% for the type Ⅱ group (6/118), and 14.3% for the type Ⅲ group (3/21), respectively. There was no significant difference among the groups ( P>0.05). The recipient cumulative survival rates at 1 and 3 years after surgery in the type I group were 97.8% and 97.0%, and the corresponding rates in the type Ⅱ group were 96.5% and 94.2%, and in the type Ⅲ group were 94.1% and 86.9%, respectively. There was a significant difference between the type Ⅰ and type Ⅲ groups ( P=0.048). The graft cumulative survival rates at 1 and 3 years in the type Ⅰ group were 97.4% and 96.9%, and the corresponding rates in the type Ⅱ group were 94.9% and 92.5%, and in the type Ⅲ group were 94.1% and 86.9%, respectively. The difference in the postoperative graft cumulative survival rates between the type Ⅰ group and type Ⅱ group was significant ( P=0.044). Conclusions:The anatomy of the left hepatic vein supplying the left lateral liver segment was highly variable, and the majority of the variations could be reconstructed. A reasonable reconstructive method could reduce the incidence of postoperative HVOO and improved the outcomes of the graft.

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.

Objective:To explore the effects of graft recipient weight ratio (GRWR) on the early prognosis (within 1 year after operation) of recipients of different ages after split liver transplantation (SLT) in children.Methods:From April 2015 to December 2022, the relevant clinical data were retrospectively reviewed for 188 children aged under 12 years undergoing initial SLT. Based upon operative age, they were assigned into groups of L (age≤18 months, 123 cases) and H (18 months< age≤12 years, 65 cases). Draw receiver operating characteristic (ROC) curves for predicting survival rates in H and L groups using GRWR and determine the cut-off value, and subgroup dassification was based the value. Compare the general condition, intraoperative condition, postoperative condition, and major complications of recipients. Follow-ups were conducted until 12 months post-SLT, death or retransplantation within 12months post-SLT. Kaplan-Meier survival rate analysis was utilized for comparing early postoperative survival rate of recipient/graft. The incidence of major early postoperative complications was examined by χ2 test or Fisher exact probability method. Results:The survival rate of recipients at Month 12 post-SLT was 92.6% (174/188), and graft survival rate was 91.0% (171/188). The survival rate of recipients in group L at Month 12 post-SLT was 94.3% (116/123), and graft survival rate was 92.7% (114/123). The GRWR value determined of 3.1 %. According to the level of GRWR, group L was divided into groups of L-L (GRWR≤3.1%, 36 cases) and L-H (GRWR>3.1%, 87 cases) while group H groups of H-L (GRWR≤3.1%, 55 cases) and H-H (GRWR>3.1%, 10 cases). The survival rates of recipients in groups L-L/L-H were 88.9% (32/36) and 96.6% (84/87) at Month 12 post-SLT. Inter-group difference was not statistically significant ( P=0.077). Graft survival rates were 83.3% (30/36) and 96.6% (84/87 ). Inter-group difference was statistically significant ( P=0.007). The intraoperative cold ischemia time were 479.0 (194.0, 593.0) min and 204.0 (122.0, 495.0) min in groups L-L/L-H. Inter-group difference was statistically significant ( P=0.002 ). The incidence of hepatic artery thrombosis were 13.9 % (5/36) and 2.3 % (2/87) in groups L-L/L-H. Inter-group difference was statistically significant ( P=0.036). The survival rate of recipients in group H at Month 12 post-SLT was 89.2% (58/65), and graft survival rate was 87.7% (57/65). No significant inter-group difference existed during surgery ( P>0.05 ). The survival rates of recipients in group H-L/H-H at Month 12 post-SLT were 92.7 % (51 /55) and 70.0 % (7/10 ). Inter-group difference was statistically significant ( P=0.019). Graft survival rates were 90.9% (50/55) and 70.0% (7/10). Inter-group difference was statistically significant ( P=0.036). No significant inter-group difference existed in the incidence of complications ( P>0.05) . Conclusion:During pediatric SLT, recipients of different ages have different requirements for GRWR. GRWR≤3.1 % implies poor early prognosis of recipients aged ≤18 months and GRWR>3.1% is associated with poor early prognosis of recipients aged between 18 months and 12 years.