Background Retinal Müller cells are important gliocytcs and the source of retinal stem cells.Researching the biological behavior of Müller cells is of important significance to the study on retinal physiopathological process and stem cell therapy of retinal diseases.To establish a stable culture method of Müller cells is a solid basis of relative basic research.Objective This study was to establish a simple and stable method of isolation and culture of human retinal Müller cells and provide sufficient and high-quality Müller cell source.Methods Human retinal Müller cells were isolated from healthy human donor eyes.The mixture solution of hyaluronidase (100 U) and 0.25％ trypsin were used to digest chopped retinal tissue.The DMEM/F12 medium with 20％ fetal bovine serum (FBS) was added to stop the digestion process.RPMI1640 medium with 20％ FBS was used to culture the cell for 72 hours and then replaced the half medium.The cells were passaged by the RPMI1640 medium with 20％ FBS.The morphology of the cells were examied under the optical microscope,and the expressions of glial fibrillary acidic protein (GFAP),a marker of gliocytes,and glutamine synthetase (GS),a special marker of retinal Müller cells,were detected by immunochemistry and immunofluorescence technology.Results Human retinal Müller cells were successfully isolated by enzyme mixture solution of hyaluronidase (100 U) and O.25％ trypsin.The cells were adherent to walls 24 hours after primary culture and completely merged 9-10 days after culture.The cells showed oval in shape with abundant cytoplasm,and a part of cells presented with cone-shaped bulge bilaterally and ectasia in the posterior containing large nuclei.After cells passage,the cells were enlarged and grew toward polygonal shape.The positive expression of GFAP was observed in more than 95％ cells and strongly positive expression of GS was observed in more than 90％ cells by immunohistochemstry and immunofluorescent staining.Conclusions Human retinal Müller cells can be successfully isolated by hyaluronidase combined with trypsin digestion.Abundent and pure human retinal Müller cells can be obtained by successively using RPMI1640 medium with 20％ FBS and 10％ FBS.

Objective:To investigate the changes and correlation of Insulin-line growth factor-1(IGF-1)and bone mineral density(BMD)in Graves disease(GD).Methods:The serum IGF-1 levels were determined by radioimmunoprecipitation assay(RIA) in 30 cases of females GD patients before and after successful therapy,the BMD were measured by dual-energy X-ray absorptiometry,the sex and ege were matched with the normal of the control group.Results:The levels of serum IGF-1(165 4?40 0) ?g/L markedly increase and the BMD(0 770?0 081)g/cm 2 markedly descend in GD group.There were significantly different from normal controls[IGF-1(92.5?25.1)?g/L、BMD(0.877?0.069)g/cm 2]( P0.05 ).Conclusion:IGF-1 probably play an important role in the pathogenesis and bone metabolism of GD.

Objective:To study the influence of the serum cytokine levels of patients with Graves′ disease(GD) before and After Treated With ~ 131 I.Methods:Using the method of radioimmunoassay, the serum levels of interleukin-1(IL-1), interleukin-2(IL-2), interleukin-6(IL-6), interleukin-8(IL-8) and tumor necrosis factor(TNF-?) of GD 52 patients were measured before and after ~ 131 I, 30 age-and sex-matched healthy subjects served as controls, and the results were compared with that of the control group.Results:The levels of IL-1, IL-6, IL-8, TNF-? of GD patients were significantly higher than that of the control group(P

Objective To observe the effects of 131I treatment on circulating granulocyte colonystimulating factor (G-CSF) and leucocyte levels of patients with Graves' disease (GD).Methods Enzyme-linked immunosorbent assay (ELISA),coulter three assortments,and radioimmunoassay were used to test the levels of circulating G-CSF,leucocytes and thyroid hormones of 65 incipient and untreated GD patients,all females,aged 21 -50,43 with normal leucocyte level and 22 with leucopenia before and after 131I treatment.Thirty age-matched healthy female subjects were used as controls.Results Before 131I treatment,the serous G-CSF level of the GD patients with normal leucocyte level was (28.4 ± 11.7)μg/L,significantly higher than that of the control [ ( 18.3 ± 6.98) μg/L,t =2.376,P ＜ 0.05 ].The serous G-CSF level of the GD patients with leucopenia was (40.1 ± 13.8 ) μg/L,significantly higher than that of the patients with normal leucocyte level ( t =2.788,P ＜ 0.01 ) and that of the control ( t =3.672,P＜0.01 ).180 d after the initiation of 131 I treatment,the G-CSF level of the patients with normal leucocyte level was (18.9 ± 8.32) μg/L,not significantly different from that of the normal controls,however,the G-CSF level of the GD patients with leucopenia was (25.7 ± 11.5) μg/L,still significantly higher than that of the normal control (t =2.103,P ＜ 0.05).The serous G-CSF level was negatively correlated with the titer of leucocyte ( r =- 0.38,P ＜ 0.05 ),however,not significantly correlated with such clinical parameters,as free triiodothyronine (FT3),free thyroxine (FT4) and thyrotropin-stimulating hormone (TSH).Conclusions Abnormal increment of G-CSF is observed in the GD patients,which may be related to the decrease of leucocyte.Effectively suppressing the auto-immune status in the GD patients,131I treatment is a safe and reliable therapy for GD patients with leucopenia and should be used as early as possible.

OBJECTIVETo investigate the effect of lovastatin on proliferation and extracellular matrix secretion of hepatic stellate cells in vitro.

METHODSRat hepatic stellate cells were incubated with different concentration of lovastatin and geranyl geranypyrophosphate. Cell proliferation was assessed by MTT colorimetric assay. Cell cycle was analysed by flow cytometry. Type IV collagen and laminin were determined by ELISA, and c-jun and c-fos expression by immunocytochemistry and computer video text analysis system.

RESULTSAddition of 0.1 to 50 micromol/L lovastatin into culture medium had no toxicity to hepatic stellate cells, but could significantly inhibit hepatic stellate cell proliferation and provoke G0/G1 phase arrest in dose-dependent manner, and could also markedly inhibit the c-jun and c-fos expression and type IV collagen and laminin secretion, which could partly be antagonized by geranyl geranypyrophosphate.

CONCLUSIONSLovastatin can significantly inhibit hepatic stellate cell proliferation and type IV collagen and laminin secretion, which might be partly related to its inhibitory effect on geranyl geranypyrophosphate formation.

Animals ; Cell Cycle ; Cell Division ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type IV ; Extracellular Matrix ; secretion ; Hepatic Stellate Cells ; drug effects ; secretion ; Lovastatin ; pharmacology ; Rats

Objective To evaluate the diagnostic performance of the histogram analysis of mono-exponential and intravoxel incoherent motion(IVIM) models to the dualistic model of epithelial ovarian cancer(EOC). Methods Forty female patients with histopathologically proven epithelial ovarian cancer underwent preoperative MR examination. Scanning sequences included conventional imaging, diffusion-weighted magnetic resonance imaging with 11 b values (0, 30, 50, 100, 150, 200, 400, 600, 800, 1 000, 1 500 s/mm2) and dynamic contrast enhanced MRI (DCE-MRI). Based on the dualistic model of EOC, all patients were divided into two groups:typeⅠ(low grade, n=16) and typeⅡ(high grade, n=24). ADC, D, D*and f maps and their corresponding histograms were generated by post-processing software. Based on an entire-tumour measurement, the following histogram parameters were recorded, respectively: (a) Mean; (b) the 10th percentile (10th);(c) the mean of the top 10 percent (MeanL);(d) the 90th percentile (90th);(e) the mean of the bottom 10 percent (MeanR). Two types were compared using independent sample t test or Mann-Whitney U test. And areas under ROC curve between two groups were assessed. Results For ADC , D, and f, all indices(Mean,10th,MeanL,90th,MeanR) of the histogram were significantly lower in typeⅡthan in type Ⅰ(P<0.05). All of parameters of D* had no significant different(P>0.05). D demonstrated a comparable accuracy with ADC in differentiating the grade of EOC (area under curve: Mean, 0.898 vs. 0.893; 10th, 0.880 vs. 0.846; MeanL, 0.878 vs. 0.858; 90th, 0.895 vs. 0.839; MeanR, 0.872 vs. 0.814), and both ADC and D have better performance than f. Conclusion It is feasible to stratify the grade of EOC by mono-exponential and IVIM models with histogram metrics,diagnositic efficiency of ADC and D values are higher.

Objective:To investigate the correlations between perfused lung volumes, visual scores (using perfusion SPECT/CT) and right-heart catheter (RHC) hemodynamic parameters in patients with chronic thromboembolic pulmonary hypertension (CTEPH).Methods:A total of 51 consecutive CTEPH patients (17 males, 34 females, age (59±12) years) in the First Affiliated Hospital of Guangzhou Medical University between March 2015 and July 2019 were retrospectively analyzed. All patients underwent lung perfusion SPECT/CT imaging and RHC examinations. Perfused lung volumes were determined using threshold-based (15%-85%) segmentation. Visual semiquantitative scoring in each lung segment was performed using Begic method. RHC hemodynamic parameters including pulmonary artery systolic pressure (PASP), pulmonary arterial diastolic pressure (PADP), mean pulmonary artery pressure (mPAP), pulmonary arteriolar wedge pressure (PAWP), pulmonary vessel resistance (PVR), cardiac output (CO), cardiac index (CI) were recorded. Spearman correlation analysis was used to evaluate the correlations between perfused lung volumes, visual scores and hemodynamic parameters.Results:There were significant correlations between perfused lung volumes (30%-70% threshold) and mPAP ( rs values: from -0.414 to -0.302, all P<0.05). Among them, perfused lung volumes under the threshold of 40% and 45% were moderately correlated with mPAP ( rs values: -0.414, -0.412, both P<0.05). Perfused lung volume (40% threshold) was moderately negatively correlated with PASP, PADP ( rs values: -0.402, -0.440, both P<0.05), and slightly negatively correlated with PVR ( rs=-0.352, P<0.05). Visual scores were slightly positively correlated with the PADP ( rs=0.311, P<0.05), while there was no correlation between visual scores and other RHC hemodynamic parameters ( rs values: from -0.201 to 0.275, all P>0.05). Conclusion:Perfused lung volumes based on threshold-based segmentation in lung perfusion SPECT/CT imaging can accurately reflect hemodynamic status and may provide useful information for severity assessment of CTEPH.

Objective: To construct a nonsuicidal selfinjury (NSSI) risk prediction model for middle school students using different machine learning algorithms and evaluate the models effectiveness, so as to provide guidance for the prevention and control of NSSI in campus. Methods: In March 2023, a total of 3 372 middle and high school students from schools in Nanchang, Fuzhou and Shangrao cities in Jiangxi Province were selected by combining stratified random cluster sampling and convenient sampling methods. Questionnaire surveys were conducted using various instruments including general information questionnaire, Selfesteem Scale, Ottawa Selfinjury Scale, Social Support Assessment Scale, Chinese Version of the Olweus Bullying Questionnaire, Event Attribution Style Scale, Adolescent Resilience Scale, and Adolescent Life Events Scale. Data were divided into training set (n=2 361) and test set (n=1 011) at a ratio of 7∶3, and variables were selected based on univariate and LASSO regression results. Four machine learning algorithms including namely random forest, support vector machine, Logistic regression and XGBoost, were used to construct NSSI risk prediction models, and the models performance was evaluated and compared using metrics including area under curve (AUC), sensitivity, specificity, positive predictive value, negative predictive value and F1 score. Results: The detection rate of NSSI among middle school students was 34.4%. Univariate analysis showed that there were statistically significant differences in NSSI detection rates among middle school students of different grades, genders, registered residence locations, whether they were class cadres and four types of bullying (physical, verbal, relational bullying and cyberbullying) (χ2=27.17, 15.81, 11.54, 4.63；68.22, 140.63, 77.81, 13.95, P<0.05). NSSI was included as the dependent variable in the LASSO regression model for variable screening, and the results regression identified 10 predictive variables including grade level, selfesteem, subjective support, support utilization, verbal bullying, emotional control, interpersonal relationships, punishment, loss of relatives and property, and health and adaptation issues. The AUC values of random forest, support vector machine, Logistic regression, and XGBoost algorithms were 0.76, 0.76, 0.76 and 0.77, respectively, with no statistically significant differences between pairwise comparisons (Z=-0.59-0.82, P>0.05). Sensitivity values were 0.62, 0.61, 0.62 and 0.61, respectively. Specificity values were 0.74, 0.78, 0.78 and 0.78, respectively. Positive predictive values were 0.56, 0.59, 0.60 and 0.59, respectively. Negative predictive values were 0.79, 0.79, 0.80 and 0.79, respectively. F1 scores were 0.59, 0.60, 0.61 and 0.60, respectively. Conclusions All four nonsuicidal selfinjury risk prediction models perform well, with the Logistic regression model slightly outperforming the others. Schools and parents should pay attention to the predictive factors corresponding to NSSI, so as to reduce the occurrence of NSSI among middle school students.

Objective To establish a novel rat model of chronic skeletal muscle composite injury in the lumbar region,to explore its mechanism and treatment method in depth.Methods Twenty-eight SD rats were randomly divided into control group(n = 10),Hitting injury model(HIM)group(n = 9),Composite damage model(CDM)group(n = 9)(vertical blow combined with forced prolonged sitting).Open field test,HE staining and PCR sequencing were performed 3 days after modeling.Results The open field distance of rats in HIM group was significantly shorter than that in the CDM group,and both groups had shorter distances than the control group.HE staining showed multiple muscle fiber breaks,significant widening of muscle gaps,uneven distribution of muscle nuclei,and visible congestion in the model group of simple blow injury;In the CDM group,muscle fibers were ruptured,the muscle gaps were slightly widened,and inflammatory cell infiltration was observed between the muscles,especially around the muscle fiber breakage,with muscle fiber nuclei gathering.A volcano plot of the sequencing result suggested significant differences in the Pf4 gene.GO analysis of muscle tissue in model group rats showed upregulation of oxygen binding and carrier activity pathways.Conclusions This study attempted to establish a composite skeletal muscle injury model to study the pathological changes of chronic skeletal muscle injury in the low back pain and evaluate the efficacy of treatment method.It has practical application value in animal experiments for chronic non-specific low back pain.

Objective To investigate the clinical characteristics of DCD donor-derived CRKP infection and bleeding in kidney transplantation,and to summarize the experience of diagnosis,treatment and prevention.Methods A retrospective analysis was carried out from July 2016 to December 2017 in hospital,containing clinical data of 4 cases of CRKP-infected DCD donors and 7 cases of kidney transplantation recipients.Results In the CRKP culture of 4 cases of DCD donors,1 case was positive for blood culture,1 case was positive for urine culture,1 case was positive for sputum culture,and 1 case was negative for blood,urine and sputum culture.The corresponding 7 recipients were all positive for blood culture after renal transplantation,4 cases were positive for urine culture,3 cases were positive for sputum culture,and 5 cases were positive for perirenal drainage.Of the 7 patients,4 cases had renal artery hemorrhage,1 of them was died.The average bleeding time was 17.75 days after operation (14-19 days).In 7 patients with renal transplantation,CRP increasd.And in 3 cases of deaths,CRP was stably higher than normal.Meanwhile,CRP in 4 surviving patients gradually decreased to the normal range after effective anti-infection treatment.All 7 patients were treated with carbapenems;2 patients were dead without avibactam therapy;and 5 cases were treated with avibactam and carbapenems and survived,1 case died and 1 case had good renal function recovery.Conclusion Positive CRKP in blood,urine and sputum of DCD donors can lead to CRKP infection in kidney transplant recipients.Even if the body fluids of donors are all negative,the false negative results could not be excluded.Persistent or increased high-level CRP after operation is an early warning on CRKP infection.And CRP can be used as an indicator for evaluating the effectiveness of anti CRKP therapy.The combination of avibactam and carbapenem antibiotics is an effective regimen in the treatment of DCD donor-derived CRKP.