Retinoblastoma (RB) is the most common intraocular malignant tumor in infants and young children. The key causative factors in the progression of RB remain unclear. Therefore, identifying genes closely associated with RB progression may provide important clues for disease diagnosis and gene therapy. However, tumor tissues have strong cellular heterogeneity. There may be significant differences in cell function and gene expression among cells in different pathological states. In this study, we downloaded single-cell transcriptome sequencing data of RB tumors and adjacent tissues from the GEO public database. Subsequently, we analyzed RB tumor transcriptional profiles with different disease duration at the single-cell level and identified cell groups and gene sets potentially associated with RB progression. The results showed that the tumor tissue and the adjacent tissues had overall consistency in the single-cell transcriptional map, but there were obvious differences in the distribution proportions of G1 phase cells, G2 phase cells, and microglia cells of cone precursors in RB tumor and the adjacent tissues. Furthermore, the role of three cell populations in the progression of RB tumors was emphatically analyzed. We found that in the early stage of RB tumors, cone precursor cells proliferated abnormally in G1 phase. With the progression of RB tumors, the proportion of cone precursor cells in G2 phase increased significantly. Meanwhile, the results of differential analysis of microglial populations during RB progression showed that the key genes mainly involved in immune response include RPL23, B2M, and HLA superfamily genes. This study provides new perspectives and data resources for the research of RB pathogenesis and progress.
Child ; Infant ; Humans ; Child, Preschool ; Retinoblastoma/pathology* ; Transcriptome ; Retinal Neoplasms/pathology*

Objective: This study aimed to analyze the influencing factors of cerebral hemodynamics index (CVHI) and provide evidence on early warning of stroke in ethnic minorities (Tujia, Buyi, Dong, Miao). Methods: From April 2017 to April 2019, ethnic minorities were examined in the Health Management Center of the Affiliated Hospital of Guizhou Medical University to determine the nationality, age, sex, medical history (such as hypertension, hyperlipidemia, and diabetes) and menstruation in women; measure physiological indexes, such as height and weight, and biochemical indexes, such as blood glucose, blood lipid, and serum uric acid levels; and conduct CVHI and bilateral arm ankle pulse wave conduction velocity (baPWV) detection. The CVHI integral was calculated and divided into normal CVHI group (≥75 points) and abnormal CVHI group (<75 points) to describe the characteristics of population distribution and analyze the related influencing factors of CVHI. Results: A total of 1 236 individuals belonging to ethnic minorities [age, 52.0±9.0 years; 575 men (46.52%) and 661 women (53.48%)]were included in the study. In ethnic minorities, 35.11% had abnormal CVHI. There was no significant difference in abnormal CVHI among ethnic groups, from highest to lowest, in Buyi nationality (38.22%), Miao nationality (37.93%), Dong nationality (32.70%), and Tujia nationality (32.36%). Minority women, menopausal patients, and patients with history of hypertension, diabetes, and hyperlipidemia more frequently had abnormal CVHI than men and non-menopausal and normal women (43.57%, 47.48%, and 57.66%, 76.19%, and 54.00% vs. 25.39%, 39.51%, and 32.30% 35.31%, and 34.74%, P<0.05). The mean BMI, systolic blood pressure (SBP), diastolic blood pressure (DBP), and FPG and TG levels, baPWV in the abnormal CVHI group were higher than those in the normal CVHI group, and height was lower the abnormal CVHI group than that in the normal group (P<0.05). The correlation analysis showed that height (r=0.309) was positively correlated with CVHI integral and SBP (r=-0.239), DBP (r=-0.189), and baPWV (r=-0.184) were negatively correlated with CVHI integral (P<0.001). The higher the height, the higher the CVHI and the lower the risk of stroke, and the higher the SBP, DBP, and baPWV, the lower the CVHI score and the higher the risk of stroke. The factor logistic regression analysis showed that old age (OR, 1.992; 95%CI, 1.405-2.825); female sex (OR, 2.240; 95%CI, 1.750-2.866); history of hypertension (OR, 3.363; 95%CI, 1.665-6.791), menopause (OR, 1.384; 95%CI, 1.016-1.885), and high SBP (OR, 1.031; 95%CI, 1.019-1.044), BMI (OR, 1.091; 95%CI, 1.048-1.135), FPG level (OR, 1.169; 95%CI, 1.017-1.344), and baPWV (OR, 1.002; 95%CI, 1.001-1.002) were risk factors for abnormal CVHI, and increased height (OR, 0.936; 95%CI 0.920-0.952) was the protective factor. Conclusion Individuals in ethnic minorities frequently have abnormal CVHI. The main risk factors of CVHI are the elderly, female, history of hypertension, menopause, systolic blood pressure, body mass index, fasting blood glucose and baPWV. It is necessary to strengthen the health management of ethnic minority gathering places, controling body weight, regularly monitoring of blood pressure and blood glucose and lipid levels, and examing cerebral vascular hemodynamics.

Objective To observe the host response and the expression of interleukin-4 (IL-4) in different cross-linked hyaluronic acid composite gels at different time points after the implantation in vivo, and to explore the significance of biocompatibility and macrophage polarization in post-implantation inflammatory response and tissue remodeling. Methods New Zealand white rabbits were respectively injected with crosslinked hyaluronic acid-crosslinked hydroxypropyl methylcellulose gel (sample 1), crosslinked hyaluronic acid-hydroxypropyl methylcellulose gel (sample 2) and commercially available modified sodium hyaluronate gel (control) in subcutaneous tissue at both sides of the spine. Then the rabbits were dissected at 1, 4 and 12 weeks after the implantation. The tissues were fixed with 10％formaldehyde solution, embedded in paraffin and sliced. Hematoxylin-eosin (HE) staining was performed to observe the degree of inflammation and fibrosis. Masson staining was performed to observe the formation of collagen fibers. Immunohistochemical staining was performed to observe the expression of IL-4. Results The results of HE staining showed that the inflammatory reaction in the sample 1 and sample 2 groups was significantly higher than that in the control group 1 and 4 weeks after the implantation. The inflammatory cells aggregated, and the wall of capsule and microcapsule was thick. The sample 1 group was more obvious, and the result was mild stimulation. For all the groups, the results were all non-irritating at 12 weeks after the implantation. The results of Masson staining showed that the collagen fibers in the sample 1 and sample 2 groups were increased compared with the control group, mainly distributed around the implantation site, and a small amount among the gels after 1 and 4 weeks. After 12 weeks, the collagen fibers were further increased, especially among the gels, which were consistent with the control group. The results of immunohistochemical staining showed that, at the same time point, the expression of IL-4 in sample 1 and sample 2 groups was higher than that in the control group, and the expression of IL-4 increased gradually with time. The expression of IL-4 in the control and sample 1 group at 12 weeks after the implantation was higher than that at 1 and 4 weeks respectively, and the differences were statistically significant (all P<0.05). In the sample 1 group, the expression of IL-4 at 12 weeks after the implantation was higher than that at 1 week, and the difference was statistically significant (P<0.05). The expression of IL-4 in the sample 1 group was significantly higher than that in the control group at 4 weeks after the implantation, and the difference was statistically significant (P<0.05). Conclusions The two different cross-linked sodium hyaluronate composite gels have good biocompatibility. The formation of collagen fiber and the expression of IL-4 can gradually increased within 12 weeks after the subcutaneous implantation, which is beneficial to the tissue remodeling.

Tetraspanins are a class of cell surface glycoproteins that are expressed very broadly, which can form a complex tetraspanins net by combining with many kinds of cell surface molecules such as integrins, signal proteins, growth factors and so on.In recent years, more and more studies suggest that tetraspanins are closely related to the invasion and metastasis of malignancies and show a certain clinical value, which can be used as new diagnosis and prognosis indicators of malignancy.

Objective To explore the clinical characteristics of neonatal dengue fever. Methods The clinical data from 4 neonates with dengue fever who were admitted and treated in 2014 were retrospectively analyzed and the related literatures had been reviewed. Results Four cases of neonatal dengue fever were all males. Three cases were mother to child transmission, the age at onset was 1 to 7 days after birth, and their mothers suffered with prenatal fever and were diagnozed of dengue fever during perinatal period. One case was community acquired, the age at onset was day 21 after birth and the neonate was bit by mosquito the day before. All four neonates had fever, two cases had rash, and one case had hemorrhagic spot. None of them had jaundice or cough. All of them had thrombocytopenia ( 30-125 )× 109/L, prolonged activated partial thromboplastin time ( 44 . 0-89 . 8 s), and increased aspartate aminotransferase (AST) ( 46-71 U/L). Three cases had declined ifbrinogen ( 1 . 36-2 . 53 g/L). Two cases had increased CK-MB ( 29-86 U/L). Two cases had increased CRP ( 3 . 00-46 . 05 mg/L). After the treatment of anti-infection and intravenous immunoglobulin, all of them were cured and discharged. The duration of hospital stay was 4-17 days. Conclusion The clinical manifestations of neonatal dengue fever were mainly fever and blood coagulation dysfunction, clinical symptoms are mild and lack of speciifcity, and prognosis are good. Mother to child transmission is one of the ways of dengue virus infection.

Objective To investigate the effects of early nasal intermittent positive pressure ventilation (NIPPV) compared with early continuous positive airway pressure (NCPAP) in low birth weight preterm infants with respiratory distress syndrome (RDS).Methods We performed a prospective,randomized controlled trial involving 364 low birth weight preterm infants with respiratory distress syndrome within 6 hours of birth.The infants were randomly assigned to NIPPV (n=188) or NCPAP (n=176) groups.Non invasive ventilation was initiated in the neonatal intensive care unit (NICU).The rate of mechanical intubation (within 3 days or 7 days),the use of pulmonary surfactant (PS),the rate of complications and mortality were compared between the two groups.Mann Whitney U,t and Chi-square tests were used for statistical analysis.Results The average time of invasive mechanical ventilation in NIPPV group were lower than that in NCPAP group[2.0 (1.0-4.0) d vs 7.0 (3.0-8.5) d,U=-3.457,P=0.001].The need for intubation and mechanical ventilation by day 3 and day 7 in the NIPPV group were less than those in the NCPAP group [day 3:4.8％ (9/188) vs 10.8％ (19/176),x2=4.621,P=0.032; day 7:9.0％ (17/188) vs 16.5％ (29/176),x2=4.551,P=0.033].In the NIPPV group,infants who got PS therapy was less than that in the NCPAP group [3.2％ (6/188) vs 8.5％ (15/176),x2=4.752,P=0.029].There was no significant difference in the fatality rate between the NIPPV and the NCPAP group [12.8％ (24/188) vs 10.8％ (19/176),P ＞ 0.05].There were no significant difference in the incidence of air leak,intracranial hemorrhage,periventricular leukomalacia,retinopathy of prematurity,necrotizing enterocolitis,patent ductus arteriosus,and bronchopulmonary dysplasia between the NIPPV group and the NCPAP group.Conclusion Among low birth weight prcterm infants with RDS,the early use of NIPPV reduces the need for PS,intubation and invasive ventilation compared with NCPAP.

Objective To explore the mechanism of rapamycin-eluted stent in vascular endothelial injury. Meth?ods (1)Rapamycin (rapamycin group) was injected to rabbit dorsum muscle to simulate rapamycin-eluted stent implanta?tion into muscles. Control group and acetone control group were established at the same time. Morphological change in mus?cle was observed and serum calcium levels were measured after rapamycin injection.(2)HUVECs were incubated with 0.1, 1, 10 and 100μg/L rapamycin for 48 h respectively or with 1μg/L rapamycin for 6, 12, 24 and 48 h respectively.Cell via?bility was examined by MTT and its relationship with drug concentration and treatment time were analyzed.(3)HUVECs were divided into control group and 1μg/L rapamycin group. After 48 h,morphological changes of HUVECs were assessed by HE staining,the production of nitric oxide was examined by Nitric Oxide Assay Kit and the intracellular calcium ion con?centration was tagged with Fluo-3/AM. Results (1) Organizational morphology in local muscle with rapamycin injection represent stent implantation of rabbit,and calcium content in local muscle increased significantly in rapamycin group com?pared with nomal control group and acetone control group(P<0.05). (2) Cell survival rate decreased significantly upon ad?ministration of rapamycin in both concentration and time dependent manner(P<0.01). (3) In rapamycin group, cytoplasm vacuoles, nucleus pycnosis and nuclear fragmentation were observed;compared with control group,the levels of intracellular free Ca2+increased while the levels of nitric oxide was reduced. Conclusion Rapamycin treatment lead of injury to vascular endothelial cells which might through up-regulating intracellular Ca2+level.

Objective To investigate the pathogenesis of small cell lung cancer ( SCLC) and to ex-plore the expression of cell cycle related kinase ( CCRK) in SCLC and its clinical significance.Methods Reverse transcription polymerase chain reaction ( RT-PCR) and Western blot were performed to examine ex-pression of CCRK in SCLC and normal tissues.Results The expressions of gene [(0.51 ±0.11)IU/L] and protein [(0.61 ±0.13)IU/L] of CCRK in SCLC tissues were significantly higher than normal tissues [(0.30 ±0.08)IU/L, (0.34 ±0.09)IU/L] ( P <0.05).The expression of CCRK was closely correlated with the clinical curative effect ( P <0.05 ) rather than the clinical stages ( P >0.05 ) .Conclusions The expressions of gene and protein of CCRK in SCLC tissues were significantly higher than normal tissues. CCRK promoted the occurrence and progress of SCLC.Chem can restrain effectually the excessive expres-sion of CCRK.The expressions of gene and protein of CCRK in the different clinical curative effect group had significant difference.

To explore the clinical application value of 3-dimensional and multiplane reconstruction with MSCT in diagnosing costal cartilage trauma. Totally 19 cases with costal cartilage trauma underwent MSCT 5 mm scanning and 0.625 mm reconstruction, and then went through three-dimensional and multiplane reconstruction. The ac-quired data were transmitted to the computer workstation through the network, and then three-reconstruction was per-formed with the software on AW4.3 platform. There were all 32 costal cartilage fractures in the 19 patients in-volving 15 cases with rib fracture, which included 2 cases and 3 fractures at the chondrosternal junction, 13 cases and 25 fractures in the middle of the costal cartilage, 4 cases and 4 fractures at the junction between costal cartilage and rib. Three-dimensional reconstruction with spiral CT could display clearly the location and number of costal cartilage frac-tures. Three-dimensional and multiplane reconstruction shows clearly the fracture and displacement of the costal cartilage, and the combination of MRP, MIR and VR may contribute to the diagnosis and clinical planning of the costal cartilage fracture.

Objective To investigate the effects of ulinastatin (UTI) combined with magnesium isoglycyrrhizinate (MgIG) on the expression of transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF) in lung tis?sue of rats with pulmonary fibrosis induced by bleomycin (BLM). Methods Ninety Wistar rats were randomly divided into five groups: BLM group, methylprednisolone (MTH) group, UTI group, MgIG group and UTI combined with MgIG (UTI+MgIG) group, n=18 for each group. The rat model of pulmonary fibrosis was established by injecting bleomycin through tra?chea in five groups. Twenty-four hours after treatment with BLM,rats were treated with normal saline every day in BLM group, and rats were treated by corresponding drugs in other groups. Six rats of each group were killed at the 7th,14th and 28th day respectively. The pathological changes of alveolitis and pulmonary fibrosis were evaluated by HE staining, and ex?pression levels of TGF-β1 and CTGF in lung tissues were detected by immunohistochemistry method. Results (1) Com?pared with BLM group, the degree of alveolitis and pulmonary fibrosis was reduced in other groups. There was significant dif?ference in alveolitis at the 7th and 14th day between UTI+MgIG group and BLM group. And there was significant difference in pulmonary fibrosis at the 14th and 28th day between UTI+MgIG group and BLM group (P<0.05). (2) Compared with BLM group, the expression levels of TGF-β1 and CTGF were decreased in other groups. In UTI+MgIG group, the expres?sion levels of TGF-β1 were significantly lower at the 7th and 14th day compared with those in UTI group and MgIG group, and which were significantly lower at the 28th day than those in MTH group, UTI group and MgIG group (P<0.05). The ex?pression levels of CTGF were significantly lower at the 7th day in UTI + MgIG group than those in UTI group and MgIG group, and which were significantly lower at the 14th and 28th day than those in MTH group, UTI group and MgIG group (P<0.05). Conclusion The combination of UTI and MgIG can alleviate alveolitis and fibrosis in BLM-induced pulmonary fibrosis rats, which might related with the down-regulation of TGF-β1 and CTGF expressions.