Objective:To observe the effects of NDRG1 on proliferation, migration and lumen formation of retinal vascular endothelial cells (RF/6A cells) in monkeys under high glucose condition. Methods:RF/6A cells were divided into normal group, mannitol group, high glucose group, small interfering RNA (siRNA) negative control group without target gene (siRNA group), 30 nmol/L siRNA down-regulated NDRG1 genome (siNDRG1 group) and 50 nmol/L siNDRG1 group. Normal group cells were cultured conventionally. The mannitol group was added with 25 mmol/L mannitol, and the high-glucose group was added with 25 mmol/L glucose. In the siRNA group, 25 mmol/L glucose was added, and then blank siRNA was added for induction. The 30 and 50 nmol/L siNDRG1 groups were added with 25 mmol/L glucose and induced with 30 and 50 nmol/L siRNDRG1, respectively. All cells were incubated for 24 h for follow-up experiments. Cell proliferation was observed by 4', 6-diaminidine 2-phenylindole staining. Cell counting kit-8 staining was used to detect cell activity. The expression level of NDRG1 mRNA and protein was detected by Western blot and real-time quantitative polymerase chain reaction. Cell migration was observed by cell scratch assay. Cell lumen formation assay was used to detect lumen formation. The two-tailed Student t test was used to compare the two groups. One-way analysis of variance was used to compare groups. Results:There were significant differences in cell proliferation rate ( t=36.659, 57.645) mobility rate ( t=24.745, 33.638) and lumen formation number ( t=41.276, 22.867) between high glucose group and normal group and mannitol group ( P <0.01). Compared with normal group and mannitol group, the relative expression levels of NDRG1 gene mRNA and protein in high glucose group were significantly decreased, with statistical significance ( t=46.145, 21.541, 36.738, 32.976; P<0.001). Compared with the siRNA negative group, the relative expression levels of NDRG1 gene mRNA and protein in 30 nmol/L siNDRG1 group and 50 nmol/L siNDRG1 group were significantly decreased, and the differences were statistically significant ( t=44.275, 40.7577, 57.167, 25.877; P<0.01). Compared with normal group and siRNA group, cell mobility in 30 nmol/LsiNDRG1 group was increased, and the difference was statistically significant ( t=57.562, 49.522; P<0.01). Compared with normal group and siRNA group, the number of cell lumen formation in 30 nmol/LsiNDRG1 group was significantly increased in the same field of vision, and the difference was statistically significant ( t=63.446, 42.742; P<0.01). Conclusion:Down-regulation of NDRG1 gene can improve the activity, migration and lumen formation of RF/6A cells under hyperglycemia.

【Objective】 Based on our previously established salt-sensitive hypertension cohort, we aimed to examine the association of genetic variants in uromodulin with blood pressure(BP) responses to dietary interventions of sodium and potassium intake. 【Methods】 In 2004, 514 subjects from 124 families in Mei County, Shaanxi Province, were recruited to establish the salt-sensitive hypertension study cohort. Among them, 333 non-parent subjects were selected and sequentially maintained on a normal-diet for 3 days, low-salt diet for 7 days, then a high-salt diet for 7 days and a high-salt diet with potassium supplementation for another 7 days. Thirteen single nucleotide polymorphisms(SNPs) in the uromodulin gene were genotyped on the MassARRAY platform. 【Results】 BP levels decreased from the baseline to low-salt diet, increased from low-salt to high-salt diet, and decreased again from the high-salt diet to the high-salt plus potassium supplementation intervention. SNPs rs77875418 and rs4997081 of the uromodulin gene were significantly associated with diastolic BP(DBP) and mean arterial pressure(MAP) responses to high-salt diet. In addition, SNPs rs77875418, rs79245268, rs4293393, rs6497476, rs4997081, rs13333226, and rs12917707 were significantly associated with systolic BP(SBP), DBP, and MAP responses to high-salt diet with potassium supplementation. 【Conclusion】 Genetic variants in uromodulin gene are significantly associated with BP responses to sodium and potassium supplementation, suggesting that uromodulin may be mechanistically involved in BP sodium-sensitivity and potassium-sensitivity.

Objective:To observe the effects of p21 activated kinase 4 (PAK4) on the mitochondrial function and biological behavior in retinal vascular endothelial cells.Methods:The experimental study was divided into two parts: in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 12 healthy C57BL/6J male mice were randomly divided into normal control group and diabetes group, with 6 mice in each group. Diabetes mice were induced by streptozotocin to establish diabetes model. Eight weeks after modeling, quantitative real-time polymerase chain reaction and Western blots were performed to detect the expression of PAK4 in diabetic retinas. In vitro cell experiments: the human retinal microvascular endothelial cells (hRMEC) were divided into three groups: conventional cultured cells group (N group), empty vector transfected (Vector group); pcDNA-PAK4 eukaryotic expression plasmid transfected group (PAK4 group). WB and qPCR were used to detect transfection efficiency, while scratching assay, cell scratch test was used to detect cell migration in hRMEC of each group. In vitro white blood cell adhesion experiment combined with 4 ', 6-diamino-2-phenylindole staining was used to detect the number of white blood cells adhering to hRMEC in each group. The Seahorse XFe96 cell energy metabolism analyzer measures intracellular mitochondrial basal respiration, adenosine triphosphate (ATP) production, maximum respiration, and reserve respiration capacity. The t-test was used for comparison between the two groups. Single factor analysis of variance was used for comparison among the three groups. Results:In vivo animal experiments: compared with normal control group, the relative expression levels of PAK4 mRNA and protein in retina of diabetic mice were significantly increased, with statistical significance ( t=25.372, 22.419, 25.372; P<0.05). In vitro cell experiment: compared with the N group and Vector group, the PAK4 protein, mRNA relative expression and cell mobility in the hRMEC of PAK4 group were significantly increased, with statistical significance ( F=36.821, 38.692, 29.421; P<0.05). Flow cytometry showed that the adhesion number of leukocytes on hRMEC in PAK4 group was significantly increased, and the difference was statistically significant ( F=39.649, P<0.01). Mitochondrial pressure measurement results showed that the capacity of mitochondrial basic respiration, ATP production, maximum respiration and reserve respiration in hRMEC in PAK4 group was significantly decreased, with statistical significance ( F=27.472, 22.315, 31.147, 27.472; P<0.05). Conclusion:Over-expression of PAK4 impairs mitochondrial function and significantly promotes leukocyte adhesion and migration in retinal vascular endothelial cells.

Objective:To observe the effect of metformin (Met) on inflammatory bodies and focal death in human retinal microvascular endothelial cells (hRMEC) in diabetes mellitus (DM) microenvironment.Methods:Experimental research was divided into in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 9 healthy C57BL/6J male mice were randomly divided into DM group, normal control group, and DM+Met group, with 3 mice in each group. DM group and DM+Met group mice were induced by streptozotocin to establish DM model, and DM+Met group was given Met 400 mg/ (kg · d) intervention. Eight weeks after modeling, the expression of NLRP3, cleaved-membrane perforating protein D (GSDMD) and cleaved-Caspase-1 in the retina of mice in the normal control group, DM group and DM+Met group were observed by immunohistochemical staining. In vitro cell experiments: hRMEC was divided into conventional culture cell group (N group), advanced glycation end products (AGE) group, and AGE+Met group. Joining the AGE, AGE+Met groups cells were induced by 150 μg/ml of glycation end products, and 2.0 mmol/L Met was added to the AGE+Met group. Pyroptosis was detected by flow cytometry; 2' ,7'-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe was used to detect the expression of reactive oxygen species (ROS) in cells of each group. Real-time fluorescence quantitative polymerase chain reaction and Western blot were used to detect the relative mRNA and protein expression levels of NLRP3, cleaved-GSDMD, cleaved-Caspase-1 in each group of cells. Single factor analysis of variance was used for comparison among the three groups.Results:In vivo animal experiments: compared with the DM group, the expression of NLRP3, cleaved-GSDMD, and cleaved-Caspase-1 in the retina of normal control group and DM+Met group mice was significantly reduced, with significant difference among the 3 groups ( F=43.478, 36.643, 24.464; P<0.01). In vitro cell experiment and flow cytometry showed that the pyroptosis rate of AGE group was significantly higher than that of N group and AGE+Met group ( F=32.598, P<0.01). The DCFH-DA detection results showed that the intracellular ROS levels in the N group and AGE+Met group were significantly lower than those in the AGE group, with the significant difference ( F=47.267, P<0.01). The mRNA ( F=51.563, 32.192, 44.473; P<0.01) and protein levels ( F=63.372, 54.463, 48.412; P<0.01) of NLRP3, cleaved-GSDMD, and cleaved-Caspase-1 in hRMEC of the AGE+Met group were significantly reduced compared to the N group. Conclusion:Met can down regulate the expression of NLRP3 inflammatory body related factors in hRMEC and inhibit pyroptosis.

Objective:To explore the impact of different dimensions of cognitive emotion regulation strategies on adolescents with unipolar depression and bipolar depression.Methods:From June 2019 to July 2021, a total of 216 adolescents with depressive disorder were selected, including 134 patients in unipolar depression group, 82 patients in bipolar depression group, and 111 normal controls were selected at the same time.Hamilton depression scale (HAMD), Hamilton anxiety scale (HAMA) and cognitive emotion regulation questionnaire (CERQ) were used to evaluate the emotional symptoms and cognitive emotion regulation strategies of all enrolled subjects. SPSS 23.0 was used for statistical analysis of the data. Kruskal-Wallis rank sum test and multiple Logistic regression analysis were used for statistical analysis.Results:There were significant differences in the dimensions of cognitive emotion regulation strategies and emotional symptoms among the three groups (all P<0.01). The scores of self-blame (14.00(12.00, 17.00), 13.50(12.00, 16.00), 12.00(11.00, 12.00)), rumination (15.00(12.00, 19.00), 14.00(12.00, 17.00), 12.00(10.00, 13.00)) and catastrophizing (13.00(11.00, 17.00), 12.00(9.00, 16.00), 8.00(6.00, 12.00)) in bipolar depression group and unipolar depression group were significantly higher than those in normal control group (all P<0.01). The score of blaming others (11.00(8.75, 13.25), 9.00(8.00, 12.00)) in bipolar depression group was significantly higher than that in normal control group ( P<0.01). The score of positive reappraisal (12.00(12.00, 15.00), 11.00(8.75, 13.00)) in normal control group was significantly higher than that in unipolar depression group ( P<0.01). The putting into perspective score(10.00(8.00, 12.00), 12.00(10.00, 13.25), 12.00(10.00, 13.00)) of normal control group was significantly lower than those of unipolar depression and bipolar depression group (both P<0.01). The scores of HAMD (25.00(22.00, 26.25), 23.00(18.00, 28.00), 3.00(0, 6.00)) and HAMA (17.00(14.00, 21.00), 20.00(16.00, 27.00), 1.00(0, 3.00)) both in unipolar depression group and bipolar depression group were significantly higher than that in normal control group (both P<0.01). Multivariate Logistic regression analysis showed that self-blame, rumination, and catastrophizing were risk factors for unipolar depression ( OR=1.19, 95% CI=1.05-1.35; OR=1.17, 95% CI= 1.06-1.30; OR=1.14, 95% CI=1.02-1.27) and bipolar depression( OR=1.30, 95% CI=1.14-1.50; OR=1.21, 95% CI=1.07-1.36; OR=1.13, 95% CI=1.01-1.28) compared to normal controls, while positive reappraisal were protective factors for unipolar depression ( OR=0.83, 95% CI=0.73-0.95) and bipolar depression ( OR=0.84, 95% CI=0.73-0.98). However, after controlling for HAMD, HAMA and gender, the effects of each dimension of cognitive emotion regulation strategies on unipolar depression and bipolar depression were no longer significant(both P>0.05). Conclusion:The negative cognitive emotion regulation strategies are correlated with the risk of disease in adolescents with unipolar and bipolar depression, and this effect is affected by the patients' own depression, anxiety and other factors.

Objective:To explore the chain mediating effect of emotion dysregulation and trait anger between childhood trauma and aggressive behavior in adolescents.Methods:A sample of 1 333 undergraduates were recruited to complete the questionnaires about childhood trauma, aggressive behavior, emotion dysregulation, trait anger.The SPSS 23. 0 and Mplus 8.3 software were used to analysis data and test intermediate effect.Results:The scores of childhood trauma, aggressive behavior, difficulties in emotion regulation and trait anger were (33.624±8.211), (53.995±12.307), (91.781±17.518), (23.352±5.477), respectively. Correlation analysis showed that childhood trauma, aggressive behavior, emotion dysregulation and trait anger were positively correlated with each other( r=0.209-0.614; all P<0.01). Mediation modeling analysis showed that childhood trauma had a significant direct effect on aggressive behavior. The direct effect value was 0.121, accounting for 35.8% of the total effect. The total indirect effect of childhood trauma on aggressive behavior was 0.217, accounting for 64.2% of the total effect. The mediating effect of emotion dysregulation as mediator between childhood trauma and aggressive behavior was 0.035, accounting for 10.4% of the total effect. The mediating effect of trait anger as mediator between childhood trauma and aggressive behavior was 0.108, accounting for 31.9% of the total effect. The chain mediating effect of emotion dysregulation and trait anger was 0.074, accounting for 21.9% of the total effect. Conclusion:Emotion dysregulation and trait anger exert a multiple mediating effect on the relationship between childhood trauma and aggressive behavior.

Objective:To investigate the short-term clinical effect of using fibular flap with preserving the continuity of fibula in hip preservation surgery for femoral head necrosis.Methods:From September, 2017 to November, 2020, 13 cases of femoral head necrosis were repaired with fibular flap. The fibular flaps were cut with an improved method for preserving the continuity of the fibular cortex, and the donor sites were sutured directly. The fibuls were inserted into the femoral heads with single or double segment folding support. Autogenous iliac crest combined with platelet-rich plasma(PRP) was used for impaction of bone grafting in femoral head, and the fibular flaps were anastomosed with 1 artery and 2 veins. All follow-up data were obtained, including bone union by X-ray and CT as well as the functional recovery of the hip joint and donor site. Statistical analysis was performed. P<0.05 was considered statistically significant. Results:The followed-up time ranged from 6 to 23 months. The fibular bones were significantly thicker and the incisions healed well at the donor sites. There was neither abnormal sensation in toes, dorsal foot, and lateral of the leg, nor significant influence on foot function. The hip joint activities were normal. The outcome was proved to be remarkable according to the Harris score(from 58.9±10.6 points before surgery to 81.7±10.6 points after surgery), the difference was statistically significant ( P<0.05) . Conclusion:The method of the improved fibular flap in hip preservation surgery is beneficial to the repair and reconstruction of the necrotic femoral head since the donor area is less traumatic, and a satisfactory clinical effect can be obtained.

Background: () (2n = 2x = 16) is genus of flowering plants belonging to the Gelsemicaeae family. Method: Here, a high-quality genome assembly using the Oxford Nanopore Technologies (ONT) platform and high-throughput chromosome conformation capture techniques (Hi-C) were used. Results: A total of 56.11 Gb of raw GridION X5 platform ONT reads (6.23 Gb per cell) were generated. After filtering, 53.45 Gb of clean reads were obtained, giving 160 × coverage depth. The genome assemblies 335.13 Mb, close to the 338 Mb estimated by k-mer analysis, was generated with contig N50 of 10.23 Mb. The vast majority (99.2%) of the assembled sequence was anchored onto 8 pseudo-chromosomes. The genome completeness was then evaluated and 1338 of the 1440 conserved genes (92.9%) could be found in the assembly. Genome annotation revealed that 43.16% of the genome is composed of repetitive elements and 23.9% is composed of long terminal repeat elements. We predicted 26,768 protein-coding genes, of which 84.56% were functionally annotated. Conclusion The genomic sequences of could be a valuable source for comparative genomic analysis in the Gelsemicaeae family and will be useful for understanding the phylogenetic relationships of the indole alkaloid metabolism.

On the basis of analysis of the current status and future tendency of development of the health services industry in Shanghai,the authors identified key problems and bottlenecks.Thus they made clear the target positioning and principles of the industry,and proposed the basic ideas and pattern of the industry in the 13 th five-year plan period,focusing on such fields as private and high-end healthcare services,traditional Chinese medicine services,public health services,commercial health insurance,and other related industries.In the end,corresponding supporting polices were proposed.

ObjectiveTo assess gene chip application value in detecting pathogenic bacteria in intracranial infection cases.MethodsPrimers and probes aiming at the specific DNA sequences of 4 kinds of common pathogenic bacteria and 6 kinds of common drug resistance genes (DRGs) were designed and used to identify the bacteria and DRGs among 30 cerebrospinal fluid (CSF) specimens (12 positive,18negative in CSF culture) from patients with intracranial infection using multiplex polymerase chain reaction (mPCR) and gene chip.The results of gene detection were compared with those of CSF culture and drug sensitivity testing.ResultsBacteria were identified and DRGs were detected in 15 specimens; DRGs and 16S gene were detected in 8 specimens; neither bacterium nor DRG was detected in 7 specimens.ConclusionGene chip technique is characterized by its relative sensitivity and rapidity of detecting the pathogenic bacteria in CSF of intraeranial infection cases.