Objective To investigate the serum expressions of vascular endothelial growth factor (VEGF), transforming growth factor-β1 (TGF-β1) and C1q/tumor necrosis factor-related protein 3 (CTRP3) in type II diabetic rats with atheroscle?rosis and to undermine the interventional mechanism of simvastatin. Methods SD rats were randomly divided into normal diet (NC) group (n=8), high-fat diet (HFD) group (n=8), high-fat diet intervention (HFD+S) group (n=8), model (M) group (n=18) and model intervention (M+S) group (n=16). The diabetic atherosclerosis model was established by streptozotocin (STZ)+Vitamin D3(VitD3)+High-fat diet. The group HFD+S and group M+S rats were administrated with simvastatin at 20 mg/(kg·d)intragastrically as intervention while distilled water [20 mL/(kg·d)] were given to other groups. Serum levels of fasting plasma glucose(FPG), blood lipid, fasting insulin(FINS), VEGF, TGF-β1 and CTRP3 were compared between each groups. Results Characteristics of atheromatous plaque were seen in group M and group M+S whose pathological change were markedly attenuated compared to group M. Serum levels of VEGF, TGF-β1 and CTRP3 were significantly high?er in rats from Group HFD than those in rats from group NC. Serum levels of VEGF and TGF-β1 were significantly higher in rats from Group M than those in rats from group NC. Serum level of VEGF was significantly higher in rats from Group M than it in rats from group HFD. Serum level of CTRP3 was significantly lower in rats from Group M than it in rats from group HFD. Moreover, serum levels of TGF-β1 and CTRP3 were significantly higher in rats from Group HFD+S than those in rats from group HFD after the intervention with simvastatin. Serum level of VEGF was significantly lower in rats from Group M+S than it in rats from group M, and serum levels of TGF-β1 and CTRP3 were significantly higher in rats from group M+S than those in rats from group M after the intervention with simvastatin. Conclusion VEGF, TGF-β1 and CTRP3 may partici?pate in development of diabetic atherosclerosis. In addition to its hypolipidemic role, Simvastatin can also down regulate se?rum level of VEGF and up regulate serum levels of TGF-β1 and CTRP3 to exert a significant protective effect on diabetic atherosclerosis.

Objective To investigate the role of GABAAα3and GABAB receptors in the ventrolateral periaqueductal gray in the development of paw acute pain in rats.Methods Twelve male SD rats, weighing 280~320 g, were randomly divided into two groups: normal saline group (group NS), formaldehyde-induced pain group (group F), 6 rats in each group.In group F, rats were subcutaneously injected with 2% formaldehyde 50 μl into the ventral surface of right hind paw to induce periphery inflammatory pain.In group NS, rats were subcutaneously injected with normal saline into the ventral surface of right hind paw.Mechanical threshold was assessed using von Frey hairs for every ten minutes.The rat pain behavior scores were recorded for every five minutes.The thickness of skin and skin temperature were recorded for every fifteen minutes.Results Mechanical hyperalgesia were induced in group F after formalin injection into right hind paw.Compared with group NS, rat pain behavior scores were increased significantly in group F at all time points after injection, mechanical threshold were decreased significantly in group F at 10-60 min after injection, the temperature of the skin and the skin thickness were increased significantly in group F at 15-60 min after injection (P<0.05), the levels of the expression of GABAAα3 and GABAB were significantly increased in group F (P<0.05).Conclusion GABAAα3and GABAB receptors mediates formalin-induced hyperalgesia at ventrolateral portion of the PAG (vlPAG) of rats.

Objective:To investigate the effects of the taurochenodeoxycholic acid(TCDCA)on cartilage degeneration in tem-poromandibular joint osteoarthritis(TMJ OA).Methods:36 female SD rats aged 6 weeks were randomly divided into 3 groups:con-trol group(CON),unilateral anterior crossbite group(UAC),UAC plus TCDCA injection group(UAC+TCDCA).UAC model was es-tablished in all rats in UAC and UAC+TCDCA groups.Samples were collected at 8 and 12 weeks(control group,UAC group,UAC+TCDCA group)after set up of the experiment(n=6),and TMJ morphological examination was performed.The expression of CYP7A1,BAAT and TGR5 in the tissue and cells was examined by immunohistochimical staining.Results:(1)Compared with the CON group of the same age,the cells in the condylar cartilage were disordered,the cartilage matrix was reduced and thinner in UAC group.Compared with UAC group of the same age,cell arrangement,cell number,cartilage matrix and cartilage thickness were im-proved in UAC+TCDCA group(P＜0.05).(2)Compared with the CON group of the same age,the positive cells for TCDCA-specific receptor TGR5 and the key enzymes CYP7A1 and BAAT were mainly distributed in the anterior hypertrophic layer and hypertrophic layer at each time point.The number of positive cells in the UAC group was significantly reduced compared with the CON group.Conclusion:TCDCA has obvious therapeutic effects on the degeneration in TMJ OA.

Oral squamous cell carcinoma (OSCC) is a common cancer that develops from oral epithelial cells, it has a high incidence, mortality and teratogenic rate, which poses a serious threat to people′s life and health.The Hippo signaling pathway plays a key role in tumorigenesis, regulation of stem cell homeostasis, tissue regeneration, and organ size control. In OSCC, activation of Hippo signaling pathway can inhibit malignant biological behavior, epithelial mesenchymal transformation and distant metastasis of tumors, and improve the survival rate of patients. Considering the importance of the Hippo signaling pathway in the development of cancer, this paper summarized the composition and regulatory mechanism of Hippo pathway, elaborated the role of Hippo signaling pathway in the occurrence and development of OSCC.At the same time, make a simple generalization about the potential therapeutic approaches and strategies to reduce the risk of drug resistance for OSCC patients targeting this pathway.

Objective:To explore the research hotspots of early enteral nutrition and analyze its development trend.Methods:The Web of Science core database was retrieved. HistCite and CiteSpace were used to conduct quantitative analysis and co-word clustering analysis of early enteral nutrition.Results:A total of 823 articles were retrieved, and the number of articles was increasing. The research hotspots of early enteral nutrition mainly included severe disease, esophageal cancer, acute pancreatitis, sepsis, malnourished patients and premature infants. At the same time, the selection of early enteral nutrition nutrients was also a research hotspot.Conclusions:Early enteral nutrition research in critically ill patients is mature, and other specialized fields can carry out specialized early enteral nutrition support based on the research on critically ill patients. In the future, comparative studies on the effects of different nutrients in early enteral nutrition can also be carried out.

To investigate the presence of integrated Epstein-Barr virus (EBV) DNA in the NK/T cell lymphoma (NKTCL) ge-nome and analyze the integration information in the genome of NKTCL cell lines. Methods: PCR and in situ hybridization were used to detect EBV infection in five EBV (+) NK/T samples and four EBV (-) NK/T samples provided by the biobanks of the First Affiliated Hospi-tal of Zhengzhou University. Whole-genome DNA of the samples was sequenced and subjected to bioinformatics analysis. Whole-ge-nome sequence alignment was used to identify the EBV integration sequence. BLAST analysis was used to compare EBV fasta files of the samples and EBV fasta library. CREST software was used to extract softclip reads, filter all paired reads, and enumerate their distri-bution on chromosomes. The integrated genomics viewer (IGV) was used to compare the distribution of reads in partial regions of chromosome. PCR was used to amplify the high-frequency integration region of the EBV DNA. The amplified fragments were sanger se-quenced. Results: EBV DNA and EBER expression were detected in five EBV (+) NK/T samples but not in the four EBV (-) NK/T samples. Sequencing depth, coverage depth, proportion of coverage, and proportion of alignment all met the requirements for subsequent re-search. Sequence alignment revealed that the captured sequences were viral sequences. Filtered reads were most numerous in EBV (+) NKTCL cell line SNK, YTS, and EBV (+) nasal NKTCL tissue. The reads were non-randomly enriched in chromosome 2. EBV DNA inte-gration in the 400 bp region of chr2:30234084-30234483 caused insertion or deletion in the chr2p23.1 site. Conclusions: EBV DNA is highly integrated in the chr2p23.1 site of EBV (+) NKTCL cells and may affect the expression of related genes.

Peripheral nerve injury (PNI) is a common disease in the oral cavity that can easily lead to loss of function and abnormal appearance. The application of dental pulp stem cells (DPSCs) combined with tissue engineering in the repair of PNI is a research hotspot. DPSCs have the advantages of abundant sources, simple extraction, low immunogenicity and a high proliferation rate in vitro. They can differentiate into Schwann cells (SCs). SCs can induce autophagy and secrete key neurotrophic factors, such as nerve growth factor, brain-derived neurotrophic factor, ciliary neurotrophic factor and glial cell-derived neurotrophic factor. SCs are beneficial for the repair of nerve injury. DPSCs in different periods have differences in immune regulation, anti-inflammatory effects, expression of neural markers, angiogenesis and so on, which provide more diversified choices for nerve repair. At present, the introduction of tissue engineering provides a more controllable and improved microenvironment for DPSCs, which is conducive to the application and development of DPSCs in regenerative medicine and tissue engineering. However, there are still many problems to be solved, such as the selection of stem cells, functional link recovery, uncontrollable direction of axon regeneration, regulation of the peripheral nervous system and mechanism of repair.

Objective: To investigate the expression and clinical significance of programmed death-ligand 1 (PD-L1), programmed death-ligand 2 (PD-L2), and their receptor programmed cell death protein 1 (PD-1) in EBV-positive T/NK lymphoproliferative disease [Epstein-Barr virus-positive T/natural killer (NK)-cell lymphoproliferative disease, EBV(+)-T/NK-LPD]. Methods: The pathological paraffin-embedded tissues of 17 patients with EBV(+)-T/NK-LPD from the First Affiliated Hospital of Zhengzhou University from January 2013 to December 2017 were collected. These patients include 12 males and 5 females, aged 10-82 years old, the average age being 29 years, 4 people in gradeⅠ, 7 in gradeⅡ, 3 in gradeⅢ, and 3 people with hydroa vacciniforme-like lymphoproliferative disorders. Immunohistochemical SP method was used to detect the expression of PD-1, PD-L1, and PD-L2 in human EBV(+)-T/NK-LPD tissues. The relationship between PD-1, PD-L1, PD-L2 expression, and clinicopathological parameters, pathological grades and prognosis were analyzed by Fisher's exact probabilities and Spearman rank correlation. Result: After statistical analysis, the results showed that in 17 cases of tissue samples, there were 12 cases with positive PD-1 expression, 6 cases with positive PD-L1 expression and 5 cases with positive PD-L2 expression. There was no significant correlation between PD-1 and PD-L2 expression and prognosis (P>0.05). PD-L1 expression showed a positive correlation with prognosis (P<0.05). There was no significant correlation between the expression of PD-L1 and PD-L2 with age, sex, as well as LDH and Ki-67 levels (P>0.05). Moreover, there was no significant correlation of PD-1 and PD-L2 expression with pathological grade (r=0.141, r=-0.149, both P>0.05). However, there was a negative correlation between the PD-L1 expression and pathological grade (r=-0.563), and the correlation between the PD-L1 ex-pression and pathological grade was statistically significant (P<0.05). Conclusions: PD-1, PD-L1, and PD-L2 are abnormally expressed in the pathological tissues of EBV(+)-T/NK-LPD. Although there was no significant correlation between the expression of PD-1 and prognosis or pathological grade, it was significantly higher in EBV+T/NK-LPD. PD-1/PD-Ls associated signaling pathway is expected to be a potential new target for EBV(+)-T/NK-LPD immunotherapy.

Objective : To investigate the impact of SOX4 on ovarian granulosa cells，stable overexpression of SOX4 was achieved in human KGN cell line，followed by analysis of its effects on proliferation，migration and apoptosis. Methods : The recombinant lentiviral plasmid pLV-EF1a-GFP / Puro-SOX4 was generated through homologous recombination with linearized pLV-EF1a-GFP / Puro vector．Human ovarian granulosa cells ( KGN cell line ) were transduced with Lentiviral expression vectors．KGN cells infected with pLV-EF1a-GFP / Puro-NC were served as the LV-CON group，while those infected with pLV-EF1a-GFP / Puro-SOX4 were designated as the LV-SOX4 group．Following transfection，puromycin selection was employed to establish stable SOX4-expressing KGN cells．The expres- sion levels of SOX4 m RNA and protein in KGN cells from the LV-CON and LV-SOX4 groups were assessed using RT-qPCR and Western blot analysis．Cell proliferation was assessed using the CCK-8 assay in both LV-CON and LV-SOX4 groups．Cell migration ability was evaluated by means of a cell scratch test in these two groups．The proportion of apoptotic cells was determined via flow cytometry analysis in both LV-CON and LV-SOX4 groups. Results: The sequencing results of pLV-EF1a-GFP / Puro-SOX4 indicated a complete match between the inserted gene se- quence and the SOX4 mRNA sequence．The lentiviral titers were 7 × 108 TU / ml in the LV-CON group and 1 × 108 TU / ml in the LV-SOX4 group．The recombinant plasmid was successfully transfected into KGN cells with a transfection efficiency of over 90% under fluorescence inverted microscopy．The results of RT-qPCR and Western blot tests demonstrated a significant increase in the expression level of SOX4 in KGN cells of LV-SOX4 group compared to that of LV-CON group (t = 3. 10，P ＜0. 05 ; t = 14. 20，P ＜0. 05) ．The CCK-8 assay results demonstrated that the LV-SOX4 group exhibited a significant increase in cell proliferation (24 h : t = 45. 92，P＜0. 01 ; 72 h : t = 25. 60，P ＜0. 01) compared to the LV-CON group．The cell scratch assay indicated that the migratory capacity of KGN cells in the LV-SOX4 group was significantly enhanced (t = 7. 65，P ＜0. 01) compared to that in the LV-CON group. The LV-SOX4 group exhibited a significant reduction in apoptosis ratio (t = 25. 84，P＜0. 01) compared to the LV- CON group． Conclusion SOX4-overexpressing KGN cell line was successfully established，and the overexpression of SOX4 facilitated proliferation and migration while inhibiting apoptosis in human ovarian granulosa cells.

Objective: To investigate the effect of neurite outgrowth inhibitor extracellular peptide residues 1-40 (NEP1-40) combined with poly (lactic-co-glycolic acid) (PLGA) and gelatin electrospun fiber membrane on facial nerve repair in rats. Methods: According to the principle of random grouping, 108 male SD rats were divided into four groups (n = 27 in each group, approved by the ethics committee), namely, the sham group, control group, PLGA group, and NEP1-40 + PLGA group. A facial nerve fracture model was established for all of the groups except for the sham group. The control group received no further treatment, the PLGA group and the NEP1-40+PLGA group were supported by PLGA membrane, and the NEP1-40+PLGA group received one immediate local injection of NEP1-40 (5 μg/μL) at a dose of 10 μL. Facial nerve function analysis, electrophysiological examination, transmission electron microscope observation, HE staining, and immunohistochemical staining of myelin marker S100β and axonal marker β3-tubulin were used to evaluate the recovery of injured facial nerves of rats at 2, 4 and 8 weeks. Results : At 8 weeks, the facial nerve function score of the NEP1-40+PLGA group was better than that of the control group and PLGA group (P < 0.001), and facial nerve function was significantly restored. Electrophysiological examination of nerve action potentials at the injured facial nerve showed that the amplitude in the NEP1-40+PLGA group was higher than that of the control group and PLGA group (P < 0.001), but there was no significant difference in latency and conduction velocity results between the groups (P > 0.05). At 2, 4, and 8 weeks, transmission electron microscopy showed that the number of myelinated nerve fibers and myelin sheath thickness in the cross-section of the injured facial nerve in the NEP1-40+PLGA group were greater than those in the other groups (P < 0.05). At 8 weeks, HE staining showed that the facial nerves in the control group had partially recovered, but the overall cell distribution was uneven and the boundary with surrounding tissues was slightly blurred. In contrast, the NEP1-40+PLGA group had a relatively uniform cell distribution and a clearer boundary with surrounding tissues. At 2, 4, and 8 weeks, the immunohistochemical results showed that in the cross-section of the injuried facial nerve, NEP1-40 increased the expression of neural markers S100 β and β3-tubulin, especially β3-tubulin, which was close to normal levels (P > 0.05) Conclusion NEP1-40 is beneficial for the generation of new myelin sheaths and axons at the site of injury, and it can promote the repair and regeneration of injured facial nerves to a certain extent, thus accelerating the recovery of injured nerve function.