Objective To investigate the effects of c?myc promoter binding protein 1(MBP?1)gene on the proliferation of human Saos?2 osteo?sarcoma cells in vitro. Methods Saos?2 cells were divided into three groups:blank control group(untransfected cells),negative group(cells transfected with missense sequence)and experimental group(cells transfected with MBP?1 shRNA). Two MBP?1 shRNA sequences and one neg?ative control shRNA sequence were designed ,synthesized and cloned into pSIREN?retroQ plasma. Then the recombinant plasmids were construct?ed and transfected into human Saos?2 osteosarcoma cells by Lipofectamine 2000. The expressions of MBP?1 mRNA and protein in Saos?2 cells were detected by real?time PCR and Western blot ,respectively. The effects of altered expression of MBP?1 on cell proliferation were measured by CCK?8 cell proliferation assay. The expressions of cyclin D1 and cyclin E in Saos?2 were determined by Western blot. Results PCR and sequenc?ing results indicated that the recombinant plasmids pSIREN?retroQ was constructed. The relative expression level of MBP?1 mRNA in the MBP?1 siRNA transfection group was significantly decreased than that in blank control group(P<0.05). Compared with the blank control group,the ex?pression levels of MBP?1 protein in the experimental group also significantly decreased. The proliferation abilities of Saos?2 cells at 48,72,and 96 hours after MBP?1 siRNA transfection were significantly increased than those in the blank control group(P<0.05). Compared with the blank con?trol group,the expression levels of cyclin D1 and cyclin E protein in the experimental group also significantly increased(P<0.05). Conclusion Knockdown of the expression of MBP?1 gene promotes the proliferation of human Saos?2 osteosarcoma cells. MBP?1 gene may become the new tar?get of gene therapy for osteosarcoma.

Animals ; DNA, Fungal ; Phosphorylation ; RNA Polymerase II ; Sequence Homology, Nucleic Acid ; T-Box Domain Proteins ; genetics

Objective:To undemtand the direction and synergic relationship of the reform policy of Hierarchical Medical Treatment from perspective of urban public hospital of Guangdong.Methods:Policy analysis and the rainbow model of integrated care.Results:58 pohcy documents were included in analysis.A policy package was established based on the rainbow model of integrated care.The direction of the reform policy was clear but the synergic relationship among some policy measures was still not precise.Conclusion:The policy package was on conceptual phase.It was necessary to supplement some reform policies and measures to build synergic relationship productively.

Objective To study the effects of ribosomal protein L41 (RPL41) on the proliferation and apoptosis of human retinoblastoma Y79 cells and its underlying mechanisms.Methods Y79 cells were seeded in RPMI 1640 medium containing 10％ fetal bovine serum for passage culture.Then the cells were divided into control group,with cells left untreatment,(40 μmol · L-1,80 μmol · L 1 and 120 μmol · L-1) RPL41 treatment group according to the concentration.Next CellTiter-Glo fluorescence cell viability testing system was used to observe the viability of Y79 cells in all groups,and flow cytometry was applied to measure the cell apoptotic rate in 100 μmol · L 1 RPL41 treatment group,with Hoechst staining for the observation of nuclear morphometry of apoptotic cells,and finally,Western blot was used to determine the expression of activating transcription factor 4 (ATF4) of each group.Results Compared with the control group,the viability of Y79 cells in the 40 μmol · L-1 RPL41 treatment group was (97.9 ± 1.5) ％,with no significant difference (P =0.055);and the viability in the 80 μmol · L-1 and 120 μmol · L-1 RPL41 treatment group was (87.6 ± 1.8)％ and (63.9 ± 2.0) ％,respectively,both of which were significantly different from the control group (both P ＜ 0.05),so RPL41 inhibited the viability of Y79 cells,and 100 μmol · L-1 RPL41 promoted the apoptosis of Y79 cells,with the apoptotic rate of (17.33 ± 2.47)％.Compared with normal cells,the apoptotic cells in the 100 μmol · L 1 RPL41 treatment group showed bright color and smaller cell volume by Hoechst staining.Western blot showed that PRL41 significantly decreased the expression of ATF4 protein and the expression of ATF4 protein in the 40 μmol · L 1,80 μmol · L-1 and 120 μmol · L 1 treatment group were 0.76 ± 0.04,0.29 ± 0.04,0.29 ± 0.05,respectively,all of which were significantly different from the control group (all P ＜ 0.01).Conclusion RPL41 can inhibit the proliferation and promote the apoptosis of human retinoblastoma Y79 cell,and its mechanism may be related to the expression of ATF4.

OBJECTIVETo investigate the relationship between methylation of the CDH1 gene promoter on the expression of E-cadherin and β-catenin, and to evaluate the correlation with clinicopathological characteristics of the colonic carcinoma.

METHODSMethylation specific PCR (MSP) was used to detect CDH1 gene promoter methylation in the cancer tissue, adjacent tissues and normal tissues in 68 patients. The expression of E-cadherin and β-catenin was determined by immunohistochemistry staining.

RESULTSThe positive rate of CDH1 gene promoter methylation was 32.4% in adjacent tissues and 57.4% in cancer tissue, while no detectable methylation was found in all the normal tissues. The difference was statistically significant. The positive rate of E-cadherin was 92.6% in the normal tissues, 66.2% in the adjacent tissues and 44.1% in the cancer tissues. In all normal tissues, β-catenin was expressed only at the cellular membrane but not in the cytosol or nucleus, while the expression of β-catenin was present in the cytosol or nucleus in 29.4% of the adjacent tissues and 50.0% of the cancer tissues. The positive rate of CDH1 gene promoter methylation was negatively correlated with E-cadherin expression(r=-0.312, P=0.01) and positively correlated with β-catenin cytosolic/nucleus expression(r=0.309, P=0.018). The differentiation and metastasis of colonic carcinoma were associated with the aberrant expression of E-cadherin, β-catenin, and methylation of CDH1 promoter (P<0.05).

CONCLUSIONCDH1 gene promoter methylation may lead to aberrant expression of E-cadherin and β-catenin in colonic carcinoma, and may play an important role in promoting the invasion of tumor.

Adult ; Aged ; Aged, 80 and over ; Cadherins ; genetics ; metabolism ; Colonic Neoplasms ; genetics ; metabolism ; DNA Methylation ; Female ; Humans ; Male ; Middle Aged ; Promoter Regions, Genetic ; beta Catenin ; genetics ; metabolism

BACKGROUND: Chinese medicine is effective for preventing and treating glucocorticoid-induced osteoporosis, however, the underlying mechanism remains unclear. DKK1, an inhibitor of Wnt/β-catenin signaling pathway, can be up-regulated by glucocorticoid. Thereafter, DKK1 is an important target in the prevention and treatment of osteoporosis. OBJECTIVE: To explore the regulatory effect of Zuogui pill on DKK1 in the prevention and treatment of glucocorticoid-induced osteoporosis. METHODS: Eighteen three-month-old female Sprague-Dawley rats were randomly divided into three groups: control group, model group and Zuogui pill group. Rats in the model and Zuogui pill groups received the subcutaneous injection of dexamethasone to establish the model of glucocorticoid-induced osteoporosis. The Zuogui pill group rats were administrated Zuogui pill extracts, and the control rats were given the same volume of normal saline. At 1 month after modeling, the lumbar vertebrae were removed to test the bone mass and microstructures by micro-CT scanning. The biomechanical properties were detected by compression test. The mRNA expression levels of DKK1, Runx2 and CTSK were determined by Qpcr. The serum alkaline phosphatase activity was tested. RESULTS AND CONCLUSION: Compared with the control group, in the model group, the volumetric bone mineral density, trabecular bone volume fraction, trabecular number, and trabecular thickness were significantly decreased (P ＜ 0.05), the trabecular separation and structure model index were significantly increased (P ＜ 0.05). The serum alkaline phosphatase activity was on a decline. The mRNA expression level of DKK1 showed a significant up-regulation (P ＜ 0.05). The mRNA expression level of Runx2 showed a down-regulated trend while mRNA expression level of CTSK showed an up-regulated trend. Compared with the model group, the Zuogui pill group showed significantly enhanced volumetric bone mineral density, trabecular bone volume fraction, and trabecular number (P ＜ 0.05); the structure model index was significantly decreased (P ＜ 0.05); the trabecular separation was reduced; the serum alkaline phosphatase activity was enhanced; the mRNA expression level of DKK1 showed a significant down-regulation (P ＜ 0.05); the mRNA expression level of Runx2 showed an up-regulated trend while mRNA expression level of CTSK showed a down-regulated trend. The vertebral compressive strength in the Zuogui pill group was significantly higher than that in the model group (P＜0.05). In summary, Zuogui pill prevents and treats glucocorticoid-induced osteoporosis possibly through the down-regulation of mRNA expression of DKK1.

Objective To explore the correlation between the level of serum homocysteine (Hcy) and the thickness of retinal ganglion cell layer (GCL) on optical coherence tomography (OCT) in patients with type 2 diabetes.Methods Totally 60 diabetic patients were collected from October 2016 to October 2017 in the Shengjing Hospital of China Medical University,and they were divided into two groups:diabetic patients without retinopathy (NDR group,n =30) and non-proliferative diabetic retinopathy group (NPDR group,n =30) according to the ETDRS classification,and meanwhile additional 30 healthy subjects were enrolled as control group.The level of serum Hcy was detected,and the retinal GCL thickness was measure using OCT in all patients for the analysis of the correlation of serum Hcy level with the thickness of GCL.Results The serum Hcy level was (11.87 ± 2.19) nmol · L-1 in the control group,(14.87 ± 0.42)nmol · L-1 in the NDR group and (20.77 ± 2.40) nmol · L-1 in the NPDR group,which was significantly increased gradually,and the difference was statistically significant (P =0.000),but the thickness of GCL was (88.33 ± 6.36) μm,(81.73 ± 1.41) μm and (64.00 ± 12.73) μm in the three groups,accordingly,which was decreased significantly gradually,with statistically significant difference (P =0.000).Along with the progress of fundus lesions,the level of serum Hcy increased,but the thickness of GCL decreased,and there was a significant negative correlation of the serum Hcy level with the thickness of GCL in the retina by Pearson (r =-0.908,P =0.000).Conclusion The increase of serum Hcy level in diabetic patients is associated with the decrease of retinal GCL thickness,and Hcy is involved in neurodegenerative changes in patients with diabetic retinopathy.

Objective To delimit the natural infectious focus, including the distribution of wildlife,species, ecology of intermediate hosts and final host of Angiostrongylus cantonensis, as well as the routes of transmission and epidemiological characteristics and wildlife of human Angiostrongylus cantonensis, based on human diverging cases identified in Shenzhen, southern area of China. Methods Data including rate of infection and density of Angiostrongylus cantonensis among different hosts in 12 different areas in Shenzhen was collected, using microscope to inspect homogenate liquids of snails. Wild mice were captured with mouse cage to examine the adult Angiostrongylus cantonensis. Using larva isolated from wild-snails-infected rats to observe the life cycle of Angiostrongylus cantonensis. Results Wild life of Angiostrongylus cantonensis existed in the southwest part of Shenzhen with its majority intermediate hosts as Achatina fulica. The overall rate of infection was 31% in wildlife and final host was found to be Rattus andersoni, Achatina fulica which were extensively distributed in the shrub region of Shenzhen because of suitable climate,humidity and vegetation for generating the life cycle of Achatina fulica. Human infected Angiostrongylus cantonensis was mainly due to eating raw snails or vegetables contaminated by larva of Angiostrongylus cantonensis.The peak of infection was seen from April to November in Shenzhen area.Conclusion Wildlife of Angiostrongylus cantonensis existed in the southwest part of Shenzhen with major wildlife reservoir including fresh water snail and wild mouse. The existence of natural focus Angiostrongylus cantonensis was now recognized as an important source of human angiostrongliasis in Shenzhen area.

OBJECTIVETo study the association between reduced folate carrier gene (RFC1 A80G) polymorphism and the risk for child with neural tube defects (NTDs), and to provide epidemiological evidence for the existence of NTDs genetic marker.

METHODSRFC1 (A80G) genotypes were detected using RFLP-PCR for blood DNA of 104 families with NTDs-affected children and 100 control families with no history of child-affected birth defects. Case-control study and transmission/disequilibrium test(TDT) for the RFC1 genotype of NTDs pedigree were carried out.

RESULTSThe G allele frequency of children with NTDs was higher than that of controls when compared to A allele( OR = 1. 64, 95% CI :1.08-2.49). The offspring of the GG genotype were associated with a 2.56-fold increased risk of NTDs when compared to the AA genotype (OR = 2.56, 95% CI: 1.04-6.36) in our study population. There was evidence of association between G allele and the risk of parent having a child with NTDs (OR = 1.56, 95% CI: 1.07-2.28) in the TDT analysis.

CONCLUSIONOur findings indicated that there was potential association between offspring RFC1 GG genotype and the risk of NTDs, and the G allele was a possible susceptible gene marker for an increased NTDs risk in the Chinese population.

Case-Control Studies ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infant ; Male ; Membrane Transport Proteins ; genetics ; Neural Tube Defects ; genetics ; Parents ; Polymorphism, Genetic ; Reduced Folate Carrier Protein

OBJECTIVES: To study the predictive factors for glucocorticoid therapy by analyzing the association between the clinical features and treatment regimens in children with eosinophilic gastroenteritis. METHODS: A retrospective analysis was performed on the medical data of 182 children with eosinophilic gastroenteritis who were admitted to Guangzhou Women and Children's Medical Center from January 2012 to December 2020. According to whether glucocorticoids were used, these children were divided into a glucocorticoid treatment group and a control group. The two groups were compared in terms of age, history of allergy, clinical symptoms, laboratory examination results, endoscopic findings, and pathological results of gastrointestinal mucosa. A multivariate logistic regression analysis was performed for the results with statistical significance. RESULTS: Of the 182 children, 36 (19.8%) received glucocorticoid therapy. The rates of hematochezia, anemia, and mucosal ulceration/luminal stenosis under endoscopy and the mucosal eosinophil infiltration count were significantly higher in the glucocorticoid treatment group than those in the control group ( CONCLUSIONS Mucosal ulceration/luminal stenosis under endoscopy or a significant increase in the mucosal eosinophil infiltration count based on pathology suggests that glucocorticoid therapy can be considered in children with eosinophil gastroenteritis.
Child ; Enteritis/drug therapy* ; Eosinophilia/drug therapy* ; Female ; Gastritis ; Glucocorticoids/therapeutic use* ; Humans ; Retrospective Studies