Objective To identify energy metabolism-proteins of mice midbrain by using the proteomic technique,and to investi-gate the relationship between these proteins and neural diseases.Methods Two-dimensional gel electrophoresis was used to sepa-rate totally soluble proteins extracted from mice midbrain.Some protein spots on two-dimensional gel electrophoresis gels were ana-lyzed by using matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).Results 24 protein spots related with energy metabolism were separated by two-dimensional gel electrophoresis and identified by using MALDI-TOF MS successfully.Conclusion The establishment of energy metabolism-related proteins map of mice midbrain lays a foundation for the research on the involvement of these proteins in neural disease pathogenesis.

Objective To investigate the anti-tumor activity of triterpenoids from Ganoderma lucidum on different tumor cells . Methods The MTT assay was adopted to detect the in vitro inhibition effect on 5 kinds of tumor cells .The inhibiting curve was drawn ,IC50 was calculated for reflecting the compound′s cytotoxic activity .Results The in vitro experiments demonstrated that three kinds of triterpenoids compound monomer showed different degrees of inhibition effect ,in which the inhibitory effect of gano-derenic acid Y was stronger ,its IC50 on H460 lung cancer cells was 22 .4 μmol/L ,followed by 7-oxo-ganoderic acid Z2 ,its IC50 was 43 .1 μmol/L .Conclusion Ganoderenic acid Y shows a strong inhibitory activity on H 460 lung cancer cells ,7-oxo-ganoderic acid Z2 shows a certain inhibitory activity on H 460 lung cancer cells ,moreover the inhibitory activity is dose dependent .The three com-pounds of ganoderenic acid Y ,7-oxo-ganoderic acid Z2 and ganoderon B have no activity or very weak activity to the other detected cell lines .The anti-lung cancer activity of ganoderenic acid Y and 7-oxo-ganoderic acid Z2 needs to be further deeply studied .

Based on network pharmacology and in vitro assays,we conducted a collaborative investi-gation into the protective effects of ginsenosides Rg1 and Re on LPS-induced damage of porcine je-junal epithelial cells IPEC-J2.Network pharmacology was used to obtain and screen the intersec-ting targets of Rg1 and Re to alleviate intestinal barrier damage,and molecular docking technique was used to verify the predicted results of network pharmacology.The experiment included the Control group,LPS group,Rg1 group,and Re group.The effects of different concentrations of Rg1 and Re on cell survival rate,apoptosis rate,TEER value,FD4 permeability,and inflammatory fac-tors of IPEC-J2 were observed,and the effects of different concentrations of Rg1 and Re on the mRNA expression levels of apoptosis-related genes were also detected by fluorescence quantitative PCR.The results of network pharmacology showed that the prevention of intestinal barrier damage by Rg1,Re mainly involved the processes of PI3K-Akt and MAPK signaling pathways.The molec-ular docking results showed that the binding energy of Rg1 to all intersecting targets was less than 0,while that of ginsenoside Re to SRC targets only was less than 0.In vitro experiments showed that pretreatment with different concentrations of Rg1 and Re increased the survival rate and TEER value of LPS-treated IPEC-J2 to varying degrees,and reduced the apoptosis,the decrease of FD4 permeability,and the secretion of inflammatory factor TNF-α,suggesting that Re and Rg1 prevented the intestinal barrier from damage.It was shown that Re and Rg1 could effectively re-duce the effects of LPS treatment on IPEC-J2 cells.Rg1 significantly upregulated the mRNA ex-pression levels of MAPK8,MAPK10,HRAS,and significantly down-regulated the mRNA expres-sion levels of MAP2K1,PIK3CG,IL-2 and SRC;and Re significantly upregulated the mRNA ex-pression levels of MAPK8,MAPK10,HRAS,and PIK3R1,BCL2 gene mRNA expression levels.These results suggest that ginsenosides Rg1,Re and ginsenoside products containing Rg1 and Re deserve further investigation in preventing intestinal barrier damage in piglets.

Objective:To prepare the chitin/hyaluronic acid/collagen hydrogel loaded with mouse adipose-derived stem cells and to explore its effects on wound healing of full-thickness skin defects in rats.Methods:The research was an experimental research. Chitin nanofibers were prepared by acid hydrolysis and alkaline extraction method, and then mixed with hyaluronic acid and collagen to prepare chitin/hyaluronic acid/collagen hydrogels (hereinafter referred to as hydrogels). Besides, the hydrogels loaded with mouse adipose-derived stem cells were prepared. Thirty male 12-week-old guinea pigs were divided into negative control group, positive control group, and hydrogel group according to the random number table, with 10 guinea pigs in each group. Ethanol, 4-aminobenzoic acid ethyl ester, or the aforementioned prepared hydrogels without cells were topically applied on both sides of back of guinea pigs respectively for induced contact and stimulated contact, and skin edema and erythema formation were observed at 24 and 48 h after stimulated contact. Adipose-derived stem cells from mice were divided into normal control group cultured routinely and hydrogel group cultured with the aforementioned prepared hydrogels without cells. After 3 d of culture, protein expressions of platelet-derived growth factor-D (PDGF-D), insulin-like growth factor-Ⅰ (IGF-Ⅰ), and transforming growth factor β 1 (TGF-β 1) were detected by Western blotting ( n=3). Eight male 8-week-old Sprague-Dawley rats were taken and a circular full-thickness skin defect wound was created on each side of the back. The wounds were divided into blank control group without any treatment and hydrogel group with the aforementioned prepared hydrogels loaded with adipose-derived stem cells applied. Wound healing was observed at 0 (immediately), 2, 4, 8, and 10 d after injury, and the wound healing rate was calculated at 2, 4, 8, and 10 d after injury. Wound tissue samples at 10 d after injury were collected, the new tissue formation was observed by hematoxylin-eosin staining; the concentrations of interleukin-1α (IL-1α), IL-6, IL-4, and IL-10 were detected by enzyme-linked immunosorbent assay method; the expressions of CD16 and CD206 positive cells were observed by immunohistochemical staining and the percentages of positive cells were calculated. The sample numbers in animal experiment were all 8. Results:At 24 h after stimulated contact, no skin edema was observed in the three groups of guinea pigs, and only mild skin erythema was observed in 7 guinea pigs in positive control group. At 48 h after stimulated contact, skin erythema was observed in 8 guinea pigs and skin edema was observed in 4 guinea pigs in positive control group, while no obvious skin erythema or edema was observed in guinea pigs in the other two groups. After 3 d of culture, the protein expression levels of PDGF-D, IGF-I, and TGF-β 1 in adipose-derived stem cells in hydrogel group were significantly higher than those in normal control group (with t values of 12.91, 11.83, and 7.92, respectively, P<0.05). From 0 to 10 d after injury, the wound areas in both groups gradually decreased, and the wounds in hydrogel group were almost completely healed at 10 d after injury. At 4, 8, and 10 d after injury, the wound healing rates in hydrogel group were (38±4)%, (54±5)%, and (69±6)%, respectively, which were significantly higher than (21±6)%, (29±7)%, and (31±7)% in blank control group (with t values of 3.82, 3.97, and 4.05, respectively, Pvalues all <0.05). At 10 d after injury, compared with those in blank control group, the epidermis in wound in hydrogel group was more intact, and there were increases in hair follicles, blood vessels, and other skin appendages. At 10 d after injury, the concentrations of IL-1α and IL-6 in wound tissue in hydrogel group were significantly lower than those in blank control group (with tvalues of 8.21 and 7.99, respectively, P<0.05), while the concentrations of IL-4 and IL-10 were significantly higher than those in blank control group (with tvalues of 6.57 and 9.03, respectively, P<0.05). The percentage of CD16 positive cells in wound tissue in hydrogel group was significantly lower than that in blank control group ( t=8.02, P<0.05), while the percentage of CD206 positive cells was significantly higher than that in blank control group ( t=7.21, P<0.05). Conclusions:The hydrogel loaded with mouse adipose-derived stem cells is non-allergenic, can promote the secretion of growth factors in adipose-derived stem cells, promote the polarization of macrophages to M2 phenotype in wound tissue in rats with full-thickness skin defects, and alleviate inflammatory reaction, thereby promoting wound healing.

The antigen gene expression level of a DNA vaccine is the key factor influencing the efficacy of the DNA vaccine. Accordingly, one of the ways to improve the antigen gene expression level of a DNA vaccine is to utilize a plasmid vector that is replicable in eukaryotic cells. A replicative DNA vaccine vector pCMVori was constructed based on the non-replicative pcDNA3.1 and the replicon of porcine circovirus 2 (PCV2) in this study. An EGFP gene was cloned into pCMVori and the control plasmid pcDNA3.1. The two recombinant vectors were transfected into PK-15 cell, and the plasmid DNA and RNA were extracted from the transfected cells. Real-time PCR was used to determine the plasmid replication efficiency of the two plasmids using plasmid before and after Bcl Ⅰ digestion as templates, and the transcription level of the Rep gene in PCV2 replicon was detected by RT-PCR. The average fluorescence intensity of cells transfected with the two plasmids was analyzed with software Image J, and the transcription level of EGFP was determined by means of real-time RT-PCR. The results showed that the replication efficiency of pCMVori in PK-15 cells incubated for 48 h was 136%, and the transcriptions of Rep and Rep' were verified by RT-PCR. The average fluorescence intensity of the cells transfected with pCMVori-EGFP was 39.14% higher than that of pcDNA3.1-EGFP, and the transcription level of EGFP in the former was also 40% higher than that in the latter. In conclusion, the DNA vaccine vector pCMVori constructed in this study can independently replicate in eukaryotic cells. As a result, the expression level of cloned target gene was elevated, providing a basis for developing the pCMVori-based DNA vaccine.
Animals ; Swine ; Circovirus/genetics* ; Vaccines, DNA/genetics* ; Replicon/genetics* ; Genetic Vectors/genetics* ; Plasmids/genetics*

During coronavirus disease 2019 epidemic, the treatment of severe trauma has been impacted. The Consensus on emergency surgery and infection prevention and control for severe trauma patients with 2019 novel corona virus pneumonia was published online on February 12, 2020, providing a strong guidance for the emergency treatment of severe trauma and the self-protection of medical staffs in the early stage of the epidemic. With the Joint Prevention and Control Mechanism of the State Council renaming "novel coronavirus pneumonia" to "novel coronavirus infection" and the infection being managed with measures against class B infectious diseases since January 8, 2023, the consensus published in 2020 is no longer applicable to the emergency treatment of severe trauma in the new stage of epidemic prevention and control. In this context, led by the Chinese Traumatology Association, Chinese Trauma Surgeon Association, Trauma Medicine Branch of Chinese International Exchange and Promotive Association for Medical and Health Care, and Editorial Board of Chinese Journal of Traumatology, the Chinese expert consensus on emergency surgery for severe trauma and infection prevention during coronavirus disease 2019 epidemic ( version 2023) is formulated to ensure the effectiveness and safety in the treatment of severe trauma in the new stage. Based on the policy of the Joint Prevention and Control Mechanism of the State Council and by using evidence-based medical evidence as well as Delphi expert consultation and voting, 16 recommendations are put forward from the four aspects of the related definitions, infection prevention, preoperative assessment and preparation, emergency operation and postoperative management, hoping to provide a reference for severe trauma care in the new stage of the epidemic prevention and control.