Objective To investigate the effect of erythropoietin on the proliferation,differentiation,and apoptosis of the cultured neonatal porcine islet cells in vitro.Methods Neonatal porcine islet cells were separated and pured from neonatal pigs with collagenase digestion and tissue culture,and their viability and purity were tested. The neonatal porcine islet cells were divided into a control group and an experimental group.The experimental group was treated with erythropoietin but not the control group,and the insulin secretion responsiveness induced by low and high glucose stimulation in the islet was tested after 5 days. Cells were counted and the activation of amplification was determined by MTT chromatometry. The rates of cell apoptosis were observed by ethidium bromide/acridine orange (EB/AO) of fluorescent light staining and flow cytometry,and the cell cycle was analyzed by flow cytometry. The expression of bcl-2,bax,caspase-3,glucose transporter 2 (GlUT-2),and pancreatic duodenal homeobox-1 (PDX-1) mRNA was tested by RT-PCR.Results After erythropoietin was treated in the cell culture,the neonatal porcine islet cells had normal morphology,function,and reaction of insulin secretion to the glucose stimulation. Cell count showed more cells in the experimental group than in the control group (P＜0.05). MTT chromatometry showed the optical absorbance tended to increase with time,and compared with the control group,the optical absorbance was higher in the experimental group (P＜0.05),the expression of PDX-1 mRNA was slightly up-regulated (P＜0.05). The expression of GLUT-2 mRNA had no difference in the 2 groups (P=0.34). In the experimental group,the apoptisis rate was lower than that in the control group by flow cytometry and EB/AO fluoscence staining (P＜0.01),and the expression of bcl-2 mRNA was higher. Howerer bax mRNA and caspase-3 mRNA were obviously lower than those in the control group (P＜0.01).Conclusion Erythropoietin can promote the proliferation but has no effect on the function of neonatal porcine islet cells in vitro. Erythropoietin can protect neonatal porcine islet cells from apoptosis through up-regulating bcl-2 mRNA and downreguling bax and caspase-3 mRNA.

Objective To investigate the protective effect and mechanism of taurine on the cytotoxicity of iohexol on HK-2 cells. Methods HK-2 cells were exposed to iohexol at different dosage (25, 50, 100, 125 gI/L) for 6 h and at the dose of 100 gl/L for different time(2 h, 4 h, 6 h). Then taurine (3,12,24 mmol/L) was coincubated with iohexol (100 gI/L) for 6 h.Cell viability was assessed by CCK-8 assay. Cell apoptosis was determined by Hoechest 33342 flurescence stains,flow cytometry with Annexin V-FITC/PI double stains and caspase-3 activity by colorimetric assay. Bcl-2 and Bax expression were examined by Western blot. Intracellular ROS was detected by flow cytometry with fluorescent probe DCFH-DA. Results Iohexol decreased HK-2 cell viability and induced apoptosis in concentration-dependant and time-dependant manner (all P＜0.05). ROS was increased following iohexol (100 gI/L for 6 h) treatment (P＜0.05). Taurine increased cell viability and attenuated apoptosis in dose-dependant manner. The cell viability levels in taurine intervention (3,12,24 mmol/L) group were significantly increased compared with that in iohexol treated group respectively [(88.00±1.00)%, (91.33±0.58)%, (95.67±1.52) % vs (76.67±1.53)%, all P＜0.05]. Apoptosis rate by flow cytometry were decreased respectively [(8.84±1.75)%,(7.86±1.82)%, (6.30±1.50)% vs (11.98±0.39)%, all P＜0.05]. Caspase-3 activities were decreased respectively [(1.33±0.10), (1.27±0.06), (1.10±0.04) vs (1.42±0.13), all P＜0.05].Taurine up-regulated the expression of Bcl-2, and decreased the intracellular ROS (all P＜0.05).Conclusions Iohexol induces cell apoptosis and oxidative stress. Taurine attenuates direct cytotoxic effect induced by iohexol. The anti-oxidative stress effect and up-regulated Bcl-2 expression may partly account for the protection of taurine.

Objective To investigate the chemical preventive effect of matrine on ethyl nitrate nitroguanidine nitrate induced gastric cancer in rats. Methods 100 male Wistar rats were selected and randomly divided into four groups,each had 25 rats ,treated with different drugs respectively.Rats in the negative control group (group A)freely drunk water;each rat in gastric carcer model group (group B)drunk ethyl nitrate nitroguanidine nitrate (ENNG)1.5 mg/d by themselves;gastric cancer model rats of experimental group (group C)drunk matrine 150 mg/(kg · d)ethyl nitrate nitroguanidine nitrate (ENNG)150mg/(kg·d)by themselves;negative control group (group D)rats drunk matrine for injection 150 mg/(kg·d)by themselves.Rats were killed after 24 weeks,rats were observed on gastric mucosa change by naked eye and microscope ,and detected proliferating cell nucleus antigen (PCNA)of stomach tissue ,levels of serum transforming growth factorβ1 (TGF-β1 )and B cell lymphoma/leukemia-2 (Bcl-2).Results Canceration rate [64.00% (16/25)]of the rat gastric mucosa in Group B was significantly higher than that in group C [12.00% (3/25)],and the difference was statistically significant (P<0.05 );PCNA,TGF-β1 and the Bcl-2 level of rats in group C was lower than those in group B,and the difference was statistically significant (P<0.05 );after raising 24 weeks,change rate of gastric mucosa atrophy and hyperplasia in group B were significantly higher than those of group C,and the difference was statistically significant (P<0.05 );after raising 24 weeks,there was no cancerous changes on gastric mucosa in group A and D ,and the change of gastric mucosa atrophy and hyperplasia had no obvious difference,there was no statistically significant difference.Conclusion Matrine could inhibit rat gastric cancer induced by ENNG by lowering PCNA,TGF-β1 and the Bcl-2 levels,which provides evidence for the potential chemical preventive effect on human gastric cancer.

Objective To investigate the clinical charactors and treatment of IgM nephropathy with nephrotic syndrome(NS) in children.Methods Thirty-six IgM nephropathy patients hospitalized in Hunan Children's Hospital as research group(group A),from June 2005 to June 2012.One hundred and six patients with minimal change disease (MCD) as control group (group B),followed up for 1 ～ 8 years.Results (1) Hematuria at presentation of the two groups respectly 3.8％ vs 30.6％ (x2 =20.403,P ＜ 0.05).(2) Renal pathology revealed that there were 26 (72.2％)patients with minimal change disease,9 cases (72.2 ％)with moderate membranoproliferative glomerulonephritis,1 cases with focal segmental glomerulosclerosis.(3) According to renal pathology,group A patients were divided into two sub groups:mild group and moderate group.To compared with group B,the steroid-resistant incidence of the 3 group were respectly 12.3％,19.2％,77.8％ (x2 =24.369,P ＜ 0.05).There was no significant difference between control group and mild group.(4)The remission rate of proteinuria in steroid-resistant patients who combined to use mycophenolate mofetil (MMF) with the two groups were respectively 50％ and 85.7 ％ (x2 =3.60,P ＞ 0.05).Conclusion Incidence of hematuria is higher in IgM nephropathy patients and patients with renal pathology for moderate lesions have a high steroid-resistant,and need use immunosuppressive early.MMF may be a good immunosuppressive for theses patients.

Objective To explore the protective effect and mechanism of antioxidant N-acetyl-L-cysteine (NAC)on the cytotoxicity induced by iohexol in HK-2 cells. Methods The incubated HK-2 cells were divided into four groups:control group,iohexol group,NAC group,and NAC+iohexol group(pre-incubated with NAC and then co-incubated with iohexol).The cell viability was tested by CCK-8 assay;cell apoptosis was determined by Hoechst 33342 fluorescence staining and flow cytometry with Annexin V-FITC/PI double staining.Intracelluar ROS waft detected by flow cytometry with DCFH-DA fluorescence staining.The signaling transduction pathways were investigated by Western blotting and immunofluorescence staining. Results Iohexol decreased cell viability,and increased apoptosis in a dose-and time-dependent manner.In iohexol(100 gl/L,6 h)group,ROS was increased by 1.30-fold of control(P＜0.05).In NAC(5,10,15 mmol/L)+iohexol groups,the cell viability was increased by 104%,118%,130%respectively,and iohexol group was 63% (P＜0.05, respectively); apoptosis rate was decreased by 13.51%, 13.46%, 12.23% respectively, and iohexol group was 24.41% (P＜0.05, respectively); ROS was decreased by 1.05-fold, 0.93-fold, 0.86-fold respectively, and iohexol group was 1.3-fold (P＜0.05, respectively).Iohexol induced the increase of p53 phosphorylatian and activity, then up-regulation of Bax and down-regulation of Bcl-2 protein expression. Iohexol induced the release of cytochrome C from mitochondria to cytoplasm, all of which caused final activation of caspase-3. The expression levels of p53, Bax and caspase-3 were decreased, while Bcl-2 protein expression level was increased by NAC. Conclusions Iohexol induces the increase of apeptosis rate and ROS generation in HK-2 cells. NAC attenuates this iohexol-induced cytotoxicity by decreasing intracelluar ROS, which is mairdy through the intrinsic pathway.

Objective:To investigate the effect of NEMO binding domain peptide (NBDP) on lung inflammation and apoptosis in mice with acute respiratory distress syndrome (ARDS) and its mechanism.Methods:Thirty-six male BALB/c mice were divided into normal saline (NS) control group, ARDS model group, NBDP negative control group and 6, 12 and 18 μg NBDP pretreatment group by random number table method, with 6 mice in each group. ARDS mouse model was reproduced by aerosol inhalation lipopolysaccharide (LPS) 50 μL. An equivalent among of NS was inhaled in NS control group. The mice in NBDP negative control group were inhaled the materials similar to the non-functional NBDP 30 minutes before the aerosol inhalation LPS; 6, 12 and 18 μg of NBDP 50 μL were respectively inhaled in NBDP pretreatment groups. After inhalation of LPS for 6 hours, mice were sacrificed to get lung tissue and observe the degree of pathological injury and edema. Western blotting was used to detect the phosphorylation of nuclear factor-κB (NF-κB) pathway related proteins [NF-κB inhibitor (IκB) kinaseα/β(IKKα/β), IκBα and NF-κB p65; p-IKKα/β, p-IκBα, p-p65] and the expression of caspase-3 in lung tissue. The bronchoalveolar lavage fluid (BALF) was collected and the levels of inflammatory markers such as myeloperoxidase (MPO), interleukins (IL-1β, IL-8), and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA).Results:ARDS model group had severe edema and hemorrhage, alveolar structure destruction, pulmonary hemorrhage and hyaline membrane formation etc. under light microscope, consistent with the pathological characteristics of ARDS lung tissue, suggesting that the ARDS model was successfully reproduced. ELISA showed that MPO, IL-1β, IL-8 and TNF-α levels of BALF in ARDS model group were obviously higher than those in NS control group. There were no significant differences in the above inflammatory indicators between NBDP negative control group and ARDS model group. The levels of MPO, IL-1β, IL-8 and TNF-α in NBDP pretreatment groups were significantly lower than those in ARDS model group in a dose-dependent manner, especially in 18 μg NBDP, the differences were statistically significant as compared with ARDS model group [MPO (ng/L): 393.32±19.35 vs. 985.87±101.50, IL-1β (ng/L): 43.05±5.11 vs. 97.68±10.88, IL-8 (ng/L): 84.64±2.32 vs. 204.00±17.37, TNF-α (ng/L): 229.13±17.03 vs. 546.73±62.72, all P < 0.05]. Western blotting showed that p-IKKα/β, p-IκBα, p-p65 and caspase-3 protein expressions in ARDS model group were significantly higher than those in NS control group. There was no significant difference in above NF-κB pathway and apoptosis-related protein expression between the NBDP negative control group and ARDS model group. The p-IKKα/β, p-IκBα, p-p65 and caspase-3 protein expression in NBDP pretreatment groups were significantly lower than those in ARDS model group in a dose-dependent manner, especially in 18 μg NBDP, the differences were statistically significant as compared with ARDS model group [p-IKKα/β protein (p-IKKα/β/β-actin): 0.15±0.02 vs. 0.42±0.04, p-IκBα protein (p-IκBα/β-actin): 0.10±0.01 vs. 0.93±0.30, p-p65 protein (p-p65/β-actin): 0.22±0.05 vs. 1.37±0.21, all P < 0.05]. Conclusion:NBDP can inhibit inflammatory response and apoptosis in ARDS lung tissue in a dose-dependent manner, and its mechanism is associated with interference NF-κB signaling pathway transduction.

Objective:To differentiate the methamphetamine users according to the developmental stages of addictive behavior, and explore the characteristics of different stages in order to provid a theoretical reference for our clinical intervention.Methods:Take the male methamphetamine users in compulsory detoxification institute whom were admitted from September 2018 to December 2019 as research objects.All the objects were asked to complete clinical diagnosis, interview and questionnaire evaluation in one week.According to the developmental stages of addictive behavior, the subjects were divided into occasional use group ( n=51), regular use group ( n=95) and compulsive use group ( n=157).All subjects were evaluated using visual analogue scale(VAS), Barrett impulsiveness scale(BIS)and CogState scale.SPSS 20.0 software was used for statistical analysis, AVOVA or Kruskal-Wallis H test was used for group comparison.Multiple Logistic regression analysis was used to explore the factors associated with the development of addictive behavior. Results:(1)The compulsive use group had higher cumulative duration(24(8, 48), 12(4, 24), 22(10, 36)), average dose(6.6±3.8, 2.8±1.4, 4.5±3.4) and craving score(1(0, 5), 0(0, 1), 1(0, 3)) than the other two groups(all P<0.05).And individuals in compulsive use group had more previous heroin use experience(20.4%, 9.8%, 14.8%, P<0.05).The regular use group had more withdrawal times than the other two groups(1(1, 3), 1(0, 1), 1(1, 2), P<0.05).The total scores of Barrett impulsiveness scale(42.8±13.3, 34.5±13.6, 36.1±14.9) and the scores in all dimensions in the compulsive use group were significantly higher than those in the other two groups(motor impulsiveness: 37.5±15.8, 27.8±13.4, 29.2±17.8; attentional impulsiveness: 43.2±18.0, 39.4±17.0, 37.2±18.1; non-planning impulsiveness: 47.2±19.8, 38.8±18.7, 40.7±20.8; P<0.05) .In the compulsive use users, the ISL(16.50±4.87, 19.30±4.78, 18.33±4.91) and SEC(0.76±0.21, 0.89±0.22, 0.81±0.21) scores about cognitive assessment were significantly lower than other two groups(both P<0.05).(2)The results of multiple Logistic regression analysis showed that cumulative duration ( β=0.022, OR=1.022, 95% CI: 1.003-1.042), dosage( β=0.625, OR=1.869, 95% CI: 1.196-2.921), craving ( β=0.194, OR=1.214, 95% CI: 1.002-1.215), the total scores of Barrett impulse scale( β=0.036, OR=1.037, 95% CI: 1.013-1.061), scores of non-planning impulsiveness( β=0.040, OR=1.041, 95% CI: 1.004-1.038), scores of motor impulsiveness( β=0.033, OR=1.214, 95% CI: 1.001-1.068) were associated with the periodic grouping of addictions. Conclusions:Addictive behavior is a progressive process and methamphetamine users at different stages have different characteristics in substance use, impulsiveness and cognitive function.The development of addictive behavior is associated with the time, dosage and craving of substance use, as well as the personality impulsivity of users.And the compulsive users suffered more cognitive impairment than the other two groups. The methamphetamine users should be identified dynamically and targeted therapeutic intervention measures should be carried out to block the addictive process to achieve the goal of harm reduction.

Objective:To explore the clinical immune manifestations and high risk factors of children with neuropsychiatric systemic lupus erythematosus (NPSLE), so as to find the basis for early clinical differential diagnosis, prevention and treatment of NPSLE.Methods:Ninety-four children with systemic lupus erythematosus (SLE) admitted to Hunan Children′s Hospital from January 1, 2015 to April 30, 2022 were retrospectively selected. Among them, 19 children with NPSLE were in the NPSLE group, and 75 children without NPSLE were in the SLE group. The disease activity, immune system, lymphocyte subsets, inflammatory factors and autoantibodies were compared between the two groups.Results:Among 94 children with SLE, the incidence of NPSLE was 20.21%(19/94). The most common symptoms of 19 children with NPSLE were headache, epileptic seizure and cerebrovascular disease. The Systemic Lupus Erythematosus Disease Activity Index (SLEDAI)-2000 score was (19.89±8.33)points. Most of them had a good prognosis through hormone intervention. Compared with the SLE group, NPSLE group had higher SLEDAI-2000 score, white blood cell count, urine protein, erythrocyte sedimentation rate (ESR), interferon (IFN)-α, positive rate of anti-Sm antibody and anticardiolipin antibody, lower platelet count, interleukin-2 (IL-2), C 3, C 4 level (all P<0.05), while there was no significant difference in positive rate of anti-dsDNA antibody and anti-SSA antibody between the two group (both P>0.05). Conclusions:Compared with SLE children, children with NPSLE are often complicated with many kinds of abnormal immune function, mainly abnormal humoral immune function and many kinds of positive antibody, with increased inflammatory factors.

Objective To investigate the study of magnetic resonance (MR) diffusion tensor imaging in children with acute kidney injury,and further improve the clinical research level of early diagnosis of acute kidney injury (AKI).Methods Twenty-two children who met the clinical AKI diagnostic criteria were collected from the Children's Hospital of Hunan Province.Twenty-three children volunteers were collected as the control group.The anisotropy fraction (FA) and mean diffusion coefficient (ADC) values of the renal cortex and medulla of all the tested children were detected,and the serum creatinine value and disease test results of children with AKI were collected.Spearman correlation analysis was used to analyze the correlation between the renal and medullary ADC values and FA values and serum creatinine values in the children with AKI.Results There were no significant differences in the FA,ADC values of left and right renal cortex and medulla of case group (P ＞ 0.05).There were no significant differences in the FA,ADC values of left and right renal cortex and medulla of control group (P ＞ 0.05).The medullary FA value,cortical FA and ADC value of the children with AKI were significantly lower than those of normal children (P ＜ 0.05).There was no significant difference in medullary ADC values between children with AKI and normal children (P ＞ 0.05).The medullary FA value and cortical ADC value of AKI patients were negatively correlated with serum creatinine value (r =-0.868,-0.436,P ＜ 0.05),and there was no correlation between cortical FA,medullary ADC and serum creatinine in the rest of the children.Conclusions As a non-invasive imaging method,diffusion tensor imaging (DTI) can reflect the early renal damage of AKI and has potential application value for clinical diagnosis of AKI.

Objective:To explore the mediating effect of self-control between smartphone addiction and academic procrastination in medical undergraduates.Methods:A total of 640 medical college undergraduates were selected by convenient sampling method. The self-designed general information questionnaire, smartphone addiction proneness scale (SAPS), brief self-control scale (BSCS) and procrastination assessment scale-students (PASS) were conducted among the students. SPSS 25.0 was used for descriptive statistical analysis, independent sample t test, one-way ANOVA and Pearson correlation analysis. Results:Correlation analysis showed that PASS scores were positively correlated with SAPS scores ( r=0.29, P<0.01), and negatively corrected with BSCS scores ( r=-0.26, P<0.01); the SAPS scores were negatively corrected with BSCS scores ( r=-0.33, P<0.01). Mediating effect analysis showed that the mediating role of self-control between smartphone addiction and academic procrastination were significant (effect size=0.13, 95%CI=0.03-0.26), and the mediating effect accounted for 38.24%. Conclusion:Self-control played partial mediating effect between smartphone addiction and academic procrastination in medical undergraduates. In order to improve the current situation of medical undergraduate's academic procrastination, medical college educators can intervene from the perspective of smartphone addiction and self-control.