Objective To analyze the effects of miR-181a-5p overexpression on metabolites in the small intestines of mice with subcutaneous oral cancer by detecting changes in metabolites and metabolic pathways.Methods Three groups were included in study:Control group,negative control and miR-181a-5p overexpression group.To establish a subcutaneous oral cancer model in mice,variously treated cell suspensions were subcutaneously injected into the upper right of the groin in female M-NSG severely immunodeficient mice.Changes in pathology and small intestinal tissues were assessed by HE staining.Changes in mouse body weight were also assessed.Tandem orbitrap mass spectrometry and ultra-high performance liquid chromatography-tandem time-of-flight mass spectrometry,were used to examine metabolites in the small intestines.By pre-analyzing the original data and quality rating sample data,XCMS was able to assess which metabolites were different among the groups.To identify unique metabolic pathways,KEGG enrichment analysis was used.Results A total of 170 distinct metabolites were found in the small intestinal tissues of Control and NC groups.Choline metabolism,alanine,aspartate,and glutamate metabolism,GABA synaptic metabolism,glycerophospholipid metabolism,cAMP signaling route,cancer center carbon metabolism,and niacin and niacin amine metabolic pathways were important signaling pathways for metabolite enrichment.In the NC group,16 distinct metabolites with VIP values larger than 2 were found in the small intestines compared with the OE group overexpressing miR-181a-5p.Glycerin phosphorylcholine,palmitic acid,3-hydroxybutyl carnitine,and β-hydroxybutyric acid were among the metabolites that significantly varied.The primary enhanced metabolic pathway was the choline pathway.Conclusions Mouse small intestines underwent slight changes from subcutaneous oral cancer with the greatest effect on metabolites critical for energy metabolism.The choline metabolic pathway was the pathway that selected absolutely metabolites in mouse small intestines with subcutaneous grafts of oral cancer.

Objective To investigate the effects of Gli2 on the proliferation,growth,migration,and invasion of oral cancer cells(Tca8113)at the cellular level,and to clarify the molecular mechanism of how Gli2 regulation affects the migration and invasion of oral cancer cells.Methods Small interfering(si)RNA was used to inhibit Gli2 expression in Tca8113 cells.The effects of Gli2 on the proliferation,growth,migration,and invasion of Tca8113 cells were examined by CCK-8,platb cloning,and transwell chamber assay.Further qRT-PCR and Western blot assays were used to explore the mechanism of how Gli2 regulation effects the malignant proliferation and metastasis of Tca8113 cells.Results The mRNA and protein expression of Gli2 in oral cancer cells(Tca8113)increased.Interference of Gli2 expression inhibited the proliferation,growth,migration,and invasion of Tca8113 cells.Further experiments showed that interfering with Gli2 expression inhibited the mRNA and protein expression of key factors in the Hedgehog(Hh)pathway.In addition,interference of Gli2 expression significantly affected the mRNA and protein expression of key factors in epithelial mesenchymal transformation(EMT)pathways.Conclusions Gli2 is abnormally activated during oral cancer,and interference of Gli2 expression significantly inhibits the proliferation,growth,migration,and invasion of oral cancer cells.Gli2 influences the migration and invasion of oral cancer cells by regulating the Hh and EMT pathways.This study has provided a new way to elucidate the pathogenesis of oral cancer and new perspectives on the clinical treatment of oral cancer.

Immunodeficient animal models play an important role in preclinical research and are important experimental tools in modern biomedical research that are widely used in immunology,genetics,oncology,microbiology,and other research fields.Gene editing is a technology for targeted modification of biological genomes.From emergence to application,it has greatly promoted the development of biomedical research.Gene editing technology mainly includes homing endonucleases,zinc finger nucleases,transcription activator-like effector nucleases,and the CRISPR/Cas9 system.Researchers have used these technologies to establish various types of immunodeficient animal models,each with advantages and limitations.In recent years,a large number of studies have confirmed that the human immunodeficient animal model accurately simulates the functions of cancer cells,drugs,and the human immune system,better simulates human diseases,and is widely used to study human immunobiology and the potential mechanisms of complex diseases.In this article,we review the progress in the research and application of gene editing technology to the establishment of immunodeficient animal models,discuss in depth the problems and optimization strategies of gene editing technology in the preparation of immunodeficient animal models,and present its future development prospects to provide references for researchers to select and establish immunodeficient animal models.

Objectives:To explore the changes of some peripheral blood cells related to inflammation in patients with non-arteritis central retinal artery occlusion (NA-CRAO).Methods:A retrospective clinical study. From July 2019 to July 2021, a total of 218 patients with NA-CRAO hospitalized (NA-CRAO group) in Department of Ophthalmology, Xi'an People's Hospital (Xi'an Fourth Hospital) and 218 patients with routine physical examination (control group) during the same period were included in the study. There were no significant differences in age ( t=0.60), sex composition ratio ( χ2=0.83) and body mass index ( t=0.77) between the two groups ( P>0.05). 0.2 ml fasting peripheral blood was collected from the subject, and white blood cells (WBC), neutrophils (NEUT), lymphocytes (LYMPH), red blood cells (RBC), RBC distribution width (RDW), platelets (PLT), mean PLT volume (MPV), and large PLT ratio (PLCR) were detected. The NEUT/LYMPH ratio (NLR) and PLT/LYMPH ratio (PLR) were calculated. t test was used to compare measurement data between groups. Multiple logistic regression analysis was performed for blood cells with P <0.05. The receiver operating characteristic curve (ROC curve) was used to calculate the area under the curve (AUC) and 95% confidence interval (95% CI) of each inflammatory indicator, and the optimal cutoff value was determined according to the Jorden index (sensitivity+specificity-1). Results:Compared with control group, WBC, NEUT, NLR, RDW, PLR were increased in NA-CRAO group, while RBC and LYMPH were decreased, with statistical significance ( t=9.68, 12.43, 9.47, 3.64, 5.54, 5.18, 0.46; P<0.001). There was no significant difference in PLT, MPV and PLCR between the two groups ( t=0.32, 1.56, 0.84; P>0.05). Multivariate logistic regression analysis showed that NLR was a possible risk factor for the occurrence of NA-CRAO (odds ratio=2.51, 95% CI 0.780-0.859, P=0.031). ROC curve analysis showed that the AUC predicted by NLR was 0.819, the optimal critical value was 3.05, and the sensitivity and specificity were 59.2% and 92.7%, respectively. Conclusions:In peripheral blood cells of NA-CRAO patients, NEUT is significantly increased and LYMPH is decreased. NLR is a possible risk factor for NA-CRAO.

Objective:To observe the clinical and imaging features of non-arteriotic central retinal artery occlusion (NA-CRAO) with internal boundary membrane detachment (ILMD), and to analyze its relationship with visual prognosis.Methods:A retrospective clinical study. A total of 88 patients with NA-CRAO hospitalized in Department of Ophtalmology, Xi'an People's Hospital (Xi'an Fourth Hospital) from January 2014 to June 2023 were included in the study. Best corrected visual acuity (BCVA), optical coherence tomography (OCT) and fluorescein fundus angiography (FFA) were performed. The BCVA test used the international standard visual acuity chart, which was statistically converted to the logarithm of the minimum angle of resolution (logMAR) visual acuity. OCT observed the presence of ILMD and the thickening of the inner retina and the disappearance of anatomical stratification. FFA recorded arm-retinal circulation time (A-Rct) and retinal arterion-distal filling time (FT), and observed ciliary retinal artery, fluorescein retrograde filling, cotton spots, luciferin nodal filling, macular non-perfusion, capillary fluorescein leakage, optic disc strong fluorescence, choroidal background weak fluorescence and other characteristics. According to whether there was ILMD, the patients were divided into ILMD group and non-ILMD group, with 44 cases and 44 eyes respectively. The two groups received the same treatment. The follow-up time was 30 days after treatment. The clinical, FFA characteristics and BCVA before and after treatment were compared between the two groups. t-test was used for comparison between groups. Results:In ILMD group and non-ILMD group, there were 43 cases of male and 1 case of female, respectively, and the proportion of male was significantly higher than that of female. Before and after treatment, the logMAR BCVA of ILMD group and non-ILMD group were 2.35±0.42, 2.01±0.46, 1.47±0.60, 0.77±0.49, respectively. There were significant differences in logMAR BCVA between the two groups before and after treatment ( t=8.025, 12.358; P<0.001). Before treatment, A-Rct and FT in ILMD group were longer than those in non-ILMD group, and the difference was statistically significant ( t=3.052, 3.385; P<0.05). After treatment, there was no significant difference ( t=1.040, 1.447; P >0.05). The proportion of ciliary retinal artery and cotton plaque in ILMD group was lower than that in non-ILMD group. There was no significant difference in ciliary retinal artery between the two groups ( χ2=-0.961, P>0.05), but there was a significant difference in cotton wool plaque between the two groups ( χ2=-3.364, P <0.05). Compared to the non-ILMD group, The proportion of retrograde fluorescein filling in retinal artery ( χ2=-2.846), segment filling ( χ2=-3.907), macular non-perfusion ( χ2=-6.656), capillary fluorescein leakage ( χ2=-4.367), optic disc strong fluorescence ( χ2=-3.525) and choroidal background weak fluorescence ( χ2=-2.276) increased, the difference was statistically significant ( P<0.05). Conclusions:In patients with NA-CRAO, compared with those without ILMD, those with ILMD have more severe retinal ischemia and worse BCVA before and after treatment. ILMD is one of the poor prognostic markers of NA-CRAO vision.

Objective:To compare the efficacy between dual stability constructs and modified Scott techniques for treatment of symptomatic spondylolysis in active adolescents.Methods:A retrospective study was conducted to analyze the clinical date of 64 active adolescents who had been treated for symptomatic spondylolysis at Department of Orthopedic Surgery, The 900th Hospital of Joint Logistic Support Force from January 2017 to October 2021. There were 59 males and 5 females with an age of (24.9±5.2) years. Responsible vertebral bodies were L 3 in 2 cases, L 4 in 10 cases, L 5 in 47 cases, and L 4 to L 5 in 5 cases; spondylolisthesis was accompanied in 9 cases. Depending on the surgical methods, the patients were divided into a dual stability constructs (pedicle screws and laminar screws) group (observation group, 31 cases) and a modified Scott group (control group, 33 cases). The 2 groups were compared in terms of operative time, bleeding volume, postoperative drainage volume, isthmus healing rate, rate of internal fixation failure, visual analogue scale (VAS) for low back pain, Japanese Orthopaedic Association (JOA) score and the good and excellent rate by JOA at postoperative 1 month, 3 months, and the last follow-up, and the incidence of complications. Results:There was no statistically significant difference in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). All patients were followed up. The operation time in the observation group [(94.7±14.9) min] was significantly longer than that in the control group [(84.4±16.4) min] ( P=0.011), but there was no significant difference in intraoperative bleeding volume or postoperative drainage volume ( P>0.05). The healing rate of bilateral isthmi in the observation group was 93.5% (29/31), significantly higher than that in the control group [60.6% (20/33)], and the rate of internal fixation failure in the observation group (0) was significantly lower than that in the control group (12.1%, 4/33) ( P<0.05). At postoperative 1 month, 3 months, and the last follow-up, the VAS scores were significantly lower than the preoperative value in all patients while the JOA scores significantly higher ( P<0.05). At the last follow-up, in the observation group the VAS score [0 (0, 1.0)] was significantly lower than that in the control group [1(0, 2)], and the JOA score [(27.1±1.2) points] and the excellent and good rate by JOA [93.5% (29/31)] were significantly higher than those in the control group [(25.7±2.1) points and 75.8% (25/33)] ( P<0.05). In the control group, follow-ups revealed internal fixation failure in 4 cases due to the cable cutting out of the spinous processes, yielding a failure rate of 12.1%, while no internal fixation failure was observed in the observation group. Conclusions:Both dual stability constructs and modified Scott techniques can relieve the clinical symptoms of spondylolysis in active adolescents to various extents. However, dual stability constructs with pedicle screws and laminar screws may lead to a higher isthmus healing rate and better curative effects.

Objective To analyze the clinical characteristics and screen pathogens and prognosis of anti-N-methyl-D-aspartate receptor(NMDAR)encephalitis in children and so as to improve the diagnosis and treatment of anti-NMDAR encephalitis in children.Methods A retrospective and follow-up analysis of 38 cases of pediatric anti-NMDAR encephalitis was performed,who were admitted to the West China Second University Hospital,Sichuan University from May 2012 to June 2016,and their clinical features,associated pathogen screening,treatment and progno-sis were reviewed.Results Those 38 cases included 14 males and 24 females.The age ranged from 1 year and 10 months to 17 years and 6 months.The most common symptoms were psychiatric symptoms(33 cases),and dyskinesias and movement disorders(33 cases),speech disturbances(31 cases).Twenty-four cases(63.2%)received positive pathogenic screening and only one case had right ovarian mature cystic teratoma.Among them,11 cases had positive mycoplasma pneumonia immunoglobulin M(IgM)antibodies,10 cases had positive virus screening,and 1 case had toxoplasma gondii IgM antibodies positive,and 2 cases had mix infection.Symptoms of one case were relieved after anti-toxoplasma treatment without immunosuppression treatment,and another case had partial symptom relief after mycoplas-ma pneumonia treatment.Thirty-seven cases received immunosuppressant therapy(4 cases were treated with glucocor-ticoid,12 cases were treated with glucocorticoid combined with immune globulin,2 cases were treated with glucocorti-coid combined with plasma exchange,still 19 cases were treated with glucocorticoid combined with immune globulin and plasma exchange).After follow-up for median 12 months,3 cases relapsed,29 cases were fully recovered,5 cases had mild language impairment,2 cases had mild memory impairment,and 2 cases had involuntary movement.Conclusions The most common symptoms of the patients were psychiatric symptoms,dyskinesias and movement disorders,and speech disturbances.Children with anti-NMDAR encephalitis had a low incidence of tumors,while various pathogen infections including mycoplasma pneumonia,virus and toxoplasma gondii,which may trigger abnormal immune response of anti-NMDAR encephalitis.Pathogen screening needs to be done for pediatric anti-NMDAR encephalitis and anti-patho-genic treatment may be helpful for disease remission.

Objective To study the effect of exosomes ( EXs) released from high expression of miR-132-3p mesenchymal stem cells (MSCs) on hypoxia/reoxygenation (H/R) injured endothelial cell function. Methods MSCs extracted from bone marrow of C57BL/6 mice were cultured primarily. MSCmiR-132-3p was obtained from MSCs infected with lentivirus loaded with miR-132-3p vector. At the same time,MSCNC was obtained by infecting MSCs with control lentivirus loaded with scramble sequence. EXs released from MSCNCand MSCmiR-132-3pwas isolated,and MSC-EXs and MSC-EXsmiR-132-3pwere obtained respectively. The obtained EXs and H/R damaged mouse brain microvascular endothelial cells (bend3) were co-cultured. According to culture conditions,the cells were divided into normal culture group (normal cell culture),H/R group (making a H/R model),MSC-EXs group (MSC-EXs co-culture),MSC-EXsmiR-132-3p group (MSC-EXsmiR-132-3pco-culture), and MSC-EXsmiR-132-3p+ LY294002 group ( before the cells and MSC-EXsmiR-132-3pwere co-cultured,treated by adding phosphatidyl alcohol 3 kinase [ PI3K] signaling pathway blocker LY294002 [20 μmol/L]). Quantitative real-time quantitative polymerase chain reaction was used to detect the expression of miR-132-3p in MSCs,MSC-EXs,and bend3 cells. Angiogenesis kit was used to detect angiogenic ability of bend3 cells,and 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect the proliferative capacity of bend3 cells. Scratch test was used to detect the migration ability of bend3 cells. hochest33258 staining showed cell apoptosis. Western blot was used to detect the phosphorylation level of protein kinase B ( Akt) . Results Compared with the H/R group, the MSC-EXs treatment group significantly improved the angiogenesis,proliferation,migration abilities, and Akt phosphorylation level of bend 3 cell damage induced by H/R (The H/R group were 3 ± 1,0. 275 ± 0. 020,147 ± 8 μm,and 0. 89 ± 0. 12,respectively;the MSC-EXs treatment group were 8 ± 3,0. 358 ± 0. 030,218 ± 10 μm, and 1. 37 ± 0. 25 μm,respectively;all P<0. 01). Apoptosis was significantly reduced (47 ± 2% vs. 63 ± 2%,all P<0. 01). Compared with the MSC-EXs treatment group,the angiogenesis,proliferation,migration abilities,and Akt phosphorylation level of bend 3 cells in the MSC-EXsmiR-132-3ptreatment group were increased (14 ±3,0. 444 ± 0.050,357±10μm,and1.67±0.23,respectively,all P<0.01).Apoptosis was significantly reduced (34±1%,all P<0. 01) . Compared with the MSC-EXsmiR-132-3ptreatment group, cell proliferation, migration, angiogenesis abilities,and Akt phosphorylation level in the MSC-EXsmiR-132-3p+LY294002 group were significantly reduced (5 ± 2,0. 304 ± 0. 050,175 ± 8 μm and 0. 95 ± 0. 11,respectively,all P<0. 01). Conclusion MSC-EXs with high expression of miR-132-3p may improve many physiological functions of H/R-induced damaged cerebrovascular endothelial cells by activating PI3K/Akt signaling pathway.

Objective To investigate the treatment and surgical results of acute volar plate avulsion fracture of proximal interphalangeal joint.Methods From August 2013 to June 2016,data of 15 cases of acute volar plate avulsion fracture of proximal interphalangeal joint were retrospectively analyzed.There were 8 males and 7 females aged 18-43 years old with an average of 28.6 years old.There were 4 cases of index finger,6 cases of middle finger,2 cases of ring finger and 3 cases of little finger.Eight injuries resulted from falls,four from finger breaking and three from crushing.The time from injury to operation was 5-9 days,with an average of 6.7 days.The acute volar plate avulsion fracture of proximal interphalangeal joint were treated by double thread compression suture fixation,and Kirschner wires were used to obliquely fix the interphalangeal joint.After the operation,the fore arm was fixed by plaster for 3-4 weeks.After the removal of the plaster,the flexion and extension function of the distal interphalangeal joint and the metacarpophalangeal joint was performed under the guidance of the rehabilitation instructor in a state of no weight.The finger was evaluated by X-ray every 2 weeks until the fracture healing.After 5-8 weeks,the Kirschner wire was removed when the fracture line disappeared,and the active flexion and extension of the proximal interphalangeal joint was guided by the rehabilitation instructor.Resuts In this group of 15 patients,all the incisions healed at the first stage.There was no surgical complications such as skin necrosis,and needle path infection and no abnormal finger feeling or blood circulation disorder.All patients were followed up for 8-32 months (mean,21.7 months).8 months after operation,the DASH score was 1.5 to 7.0 points with an average of 4.6 points.At the latest follow-up,there was no pain,swelling and joint contracture in the proximal interphalangeal joints.The dorsal extension of joints were stable,and the degree of active joint activity was:90°±7° of the metacarpophalangeal joint,80°±6° of the proximal interphalangeal joint,78°±7° of the distal interphalangeal joint,and 248°±22° of total active activity.According to TAM system assessment criteria:there was excellent in 12 cases,good in 2 cases,fair in 1 case,and excellent rate was 93.3％(14/15).Conclusion Double thread compression suture fixation for the treatment of acute volar plate avulsion fracture of proximal interphalangeal joints could reduce the damage to the accessory structure of the joint,and reach adequate exposure in the operationfor fixing the bone block firmly.After operation,good fracture healing and good finger function could be achieved,so the double thread compression suture fixation is an effective surgical method.

Rab27a gene can participate in the secretion, transportation and exocytosis of T lymphocytes, mast cells, neutrophils, pancreatic β cells and hematopoietic cells through its encoded proteins, and participate in the development of diseases such as albinism, cancer, diabetes, inflammation and thrombosis. In addition, the secretion regulation of Rab27a to exosomes has also received much attention. The current research status of the secretion of exosome and the genetically modified exosomes of Rab27a gene in the treatment of diseases such as inflammatory injury, thrombosis and tumor were reviewed.