Objective:To investigate the effects of gene silencing inducible cyclooxygenase-2 (COX-2) combined with hyperbaric oxygen (HBO) on neuronal cell edema, apoptosis, autophagy and neural functional recovery in rats with intracerebral hemorrhage.Methods:SPF-grade adult male SD rats ( n=96) were used to establish a cerebral hemorrhage model through stereotactic injection of thrombin VII into the caudate nucleus. They were randomized (random number) into 4 groups ( n=24/group): control group, hyperbaric oxygen (HBO) group, COX-2 RNAi group and combined group (COX-2 RNAi+HBO). The siRNA plasmid targeting silencing COX-2 gene expression was constructed. After group treatment, the four rats were randomly selected from each group for testing in each category. Postoperative day 1, 7, and 14 were assessed using the modified neurological severity score (mNSS) for evaluating neurofunctional deficits. On the 7th day, the water content of the brain tissue was measured using the dry/wet weight method. The blood-brain barrier permeability was assessed using the Evans method. Annexin V and TUNEL assays were employed to assess the apoptotic rate of neural cells. The mRNA expression level of COX-2 in brain tissue was determined using the RT-PCR method. The protein expression levels of Beclin-1, COX-2, aquaporin 4 (AQP-4), B cell lymphoma/lewkmia-2 (Bcl-2), caspase-3, hypoxia-inducible factor-1α (HIF-1α) and matrix metalloprotein-2/9 (MMP-2/9) were detected by Western blot (WB). SPSS software was used for data analysis. One-way ANOVA was used for inter group comparisons and LSD- t test was used for further pairwise comparison. Results:The SD rat intracerebral hemorrhage model and plasmid construction were successfully achieved. The mNSS scores were significantly decreased in COX-2 RNAi, HBO and combined groups compared with control group on the 7th day and 14th day (all P<0.01), especially in combined group ( P<0.01). The contents of Evans blue and the water content of brain tissue of all treatment groups were significantly lower than those in control group (all P<0.05), especially in combined group ( P<0.01). The apoptotic rate of neural cells decreased in all treatment groups compared with the control group (all P<0.05), and the combined group decreased the most ( P<0.01). The mRNA expression levels of COX-2 were significantly decreased in all treatment groups compared with the control group (all P<0.01), and combined group silenced COX-2 expression most obviously ( P<0.05). The results of WB showed that the protein expression levels of Beclin-1, COX-2, AQP-4, Caspase-3, HIF-1α, MMP-2/9 were significantly lower than control group (all P<0.05), while the expression of Bcl-2 was increased in all treatment groups (all P<0.01). Among them, the combined group exhibited the most pronounced trend ( P<0.01). Conclusions:Gene silencing of COX-2 in combination with hyperbaric oxygen therapy can effectively restore neurological function in rats with cerebral hemorrhage. The mechanism may be associated with reduced blood-brain barrier permeability, alleviated brain edema, and inhibition of neuronal apoptosis and autophagy.

ObjectiveTo explore the effects of phillygenin （PHI） on the inflammation in L02 cells induced by lipopolysaccharide （LPS） and adenosine triphosphate （ATP） and the expression of purinergic 2X7 receptor （P2X7R）， NOD-like receptor family pyrin domain containing 3 （NLRP3）， and nuclear factor kappa B （NF-κB） expression. MethodIn this study， the inflammation model was induced in L02 cells by 100 μg·L^-1 LPS treatment for 24 h and 5 mmoL·L^-1 ATP treatment for 5 h. The cells in the PHI groups were cultured with PHI （100， 50， 25 mg·L^-1） for 6 h in the LPS treatment period， followed by LPS treatment for another 18 h. After ATP treatment for 5 h， the mRNA and protein expression of interleukin-1β （IL-1β）， interleukin-18 （IL-18）， P2X7R， NLRP3， Caspase-1 precursor （pro-Caspase-1）， cleaved Caspase-1， NF-κB， and NF-κB inhibitor protein α （IκBα） in L02 cells was detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. Molecular docking was used to predict whether P2X7R could bind to PHI， and DCFH-DA was employed to detect the accumulation of reactive oxygen species （ROS） in cells. P2X7R was silenced by small interfering ribonucleic acid （siRNA）， and then the mRNA expression of IL-1β， IL-18， P2X7R， NLRP3， Caspase-1， NF-κB， and IκBα was detected by Real-time PCR. ResultReal-time PCR and Western blot showed that compared with the normal group， the model group showed increased expression of IL-1β and IL-18 （P<0.05）， and compared with the model group， the PHI groups showed down-regulated IL-1β， IL-18 mRNA and protein expression （P<0.05）. Molecular docking suggested a good binding effect of PHI to P2X7R. Real-time PCR and Western blot analysis showed that the expression of P2X7R in the model group was significantly up-regulated compared with that in the normal group （P<0.05）， and compared with the model group， the PHI groups showed down-regulated mRNA and protein expression of P2X7R （P<0.05）. DCFH-DA results showed that compared with the normal group， the model group showed increased content of ROS （P<0.05）， and compared with the model group， the PHI groups decreased the accumulation of ROS （P<0.05）. As demonstrated by Real-time PCR and Western blot， compared with the normal group， the model group showed increased expression of NLRP3 inflammasome and NF-κB （P<0.05）， and compared with the model group， the PHI groups significantly decreased the mRNA and protein expression of NLRP3 and cleaved Caspase-1， and up-regulated the mRNA and protein expression of NF-κB and IκBα （P<0.05）. Real-time PCR analysis showed that compared with the results in the model group， after silencing P2X7R by siRNA， the mRNA expression of IL-1β， IL-18， P2X7R， NLRP3， Caspase-1， NF-κB， and IκBα was decreased （P<0.05）. PHI exerted the same effect， and the mRNA expression was further reduced after the combination of them. ConclusionPHI is presumed to suppress the expression of the NLRP3/NF-κB signaling pathway by down-regulating upstream P2X7R to alleviate the LPS/ATP-induced inflammation in L02 cells， suggesting that P2X7R may be the target of PHI against inflammation.

Objective: To observe the clinical efficacy of electroacupuncture (EA) in controlling myopia in children and its effect on retinal blood flow. Methods: Sixty-eight myopic children were randomly divided into an observation group and a control group, with 34 cases in each group. The control group was given auricular acupressure treatment alone, and the observation group was treated with EA once a week in addition to the treatment used in the control group. The spherical equivalent refraction (SER) and axial length (AL) were measured at baseline, and after 3 months and 6 months of treatment. Optical coherence tomography angiography was used to measure the vessel density (VD) and perfusion density (PD) in the surface layer of the retina. Results: After 3 months and 6 months of treatment, the changes in SER between the two groups were not statistically significant (P>0.05). After 3 months of treatment, the changes in AL between the two groups were not statistically significant (P>0.05); after 6 months of treatment, the change amount of AL in the observation group was smaller than that in the control group (P<0.05); after 3 months and 6 months of treatment, the changes in VD and PD in the surface layer of the retina in the observation group were significantly greater than those in the control group (P<0.01). Conclusion: EA treatment once a week for 6 months can delay the increase of AL and improve the retinal surface blood flow in myopic children.

In the study of oral orthodontics, the dental tissue models play an important role in finite element analysis results. Currently, the commonly used alveolar bone models mainly have two kinds: the uniform and the non-uniform models. The material of the uniform model was defined with the whole alveolar bone, and each mesh element has a uniform mechanical property. While the material of the elements in non-uniform model was differently determined by the Hounsfield unit (HU) value of computed tomography (CT) images where the element was located. To investigate the effects of different alveolar bone models on the biomechanical responses of periodontal ligament (PDL), a clinical patient was chosen as the research object, his mandibular canine, PDL and two kinds of alveolar bone models were constructed, and intrusive force of 1 N and moment of 2 Nmm were exerted on the canine along its root direction, respectively, which were used to analyze the hydrostatic stress and the maximal logarithmic principal strain of PDL under different loads. Research results indicated that the mechanical responses of PDL had been affected by alveolar bone models, no matter the canine translation or rotation. Compared to the uniform model, if the alveolar bone was defined as the non-uniform model, the maximal stress and strain of PDL were decreased by 13.13% and 35.57%, respectively, when the canine translation along its root direction; while the maximal stress and strain of PDL were decreased by 19.55% and 35.64%, respectively, when the canine rotation along its root direction. The uniform alveolar bone model will induce orthodontists to choose a smaller orthodontic force. The non-uniform alveolar bone model can better reflect the differences of bone characteristics in the real alveolar bone, and more conducive to obtain accurate analysis results.
Biomechanical Phenomena ; Computer Simulation ; Finite Element Analysis ; Humans ; Models, Biological ; Periodontal Ligament ; Stress, Mechanical ; Tooth Movement Techniques

[Abstract] Objective: To explore the expression of long non-coding RNA (lncRNA) GTSE1-AS1 in prostate cancer tissues and the mechanism that affects the proliferation and invasion of LNCaP cells. Methods: From November 2017 to December 2018, 68 pairs of prostate cancer tissue and para-cancerous tissue specimens were resected from prostate cancer patients at the Department of Urology of Luoyang Central Hospital Affiliated to Zhengzhou University; in addition, prostate cancer cell lines LNCaP, PC-3, C4-2B, 22Rv1, DU-145 and normal prostate follicular epithelial RWPE-1 cells were also chosen for this study. qPCR was used to detect the expression level of GTSE1-AS1 in cancer tissues and cell lines. The GTSE1-AS1 over-expression plasmid (experimental group) and negative control plasmid (control group) were respectively transfected into LNCap cells. MTT assay and Transwell chamber method were used to detect the effect of GTSE1-AS1 over-expression on the proliferation and invasion ability of LNCaP cells, respectively. The targeting relationship among GTSE1-AS1 and miR-324-3P as well as FBXW7 (F-frame/WD repeat domain protein 7) was verified by bioinformatics tools and dual-luciferin reporter gene assay. The effect of GTSE1-AS1 over-expression on downstream gene and protein expression was detected by qPCR and WB assay. Results: The expression level of GTSE1-AS1 in prostate cancer tissues was significantly lower than that in para-cancerous tissues (P<0.01), and the expression of GTSE1-AS1 in prostate cancer cell lines was significantly lower than that in RWPE-1 cells (P<0.05 or P<0.01). Over-expression of GTSE1-AS1 significantly inhibited the proliferation and invasion (P<0.05 or P<0.01) of LNCaP cells. Dual-luciferin reporter gene assay confirmed the complementary binding between GTSE1-AS1 and miR-324-3p as well as between miR-324-3p and FBXW7. Over-expression of GTSE1-AS1 significantly reduced the expression of miR-324-3p in LNCaP cells (P<0.01), and promoted the mRNA and protein expressions of FBXW7 (all P<0.01). Conclusion: GTSE1-AS1 is under-expressed in prostate cancer tissues and cell lines. Over-expression of GTSE1-AS1 can inhibit the proliferation and invasion of LNCaP cells, the mechanism of which may be related with the inhibition of miR-324-3p to further promote FBXW7 expression.

AIMS: To investigate the association of four single-nucleotide polymorphisms (SNPs) of the IL-6 gene with osteoporosis (OST) susceptibility. METHODS: PCR restriction fragment length polymorphism (PCR-RFLP) was carried out for SNPs detection. Generalized multifactor dimensionality reduction (GMDR) model and logistic regression model were used to examine the interaction between SNP and obesity on OST. RESULTS: Logistic regression model revealed that G allele of rs1800796 and the T allele of rs2069849 were associated with increased OST risk, compared to those with wild genotype. However, no significant correlations were found when analyzing the association of rs1800795 and rs1554606 with OST risk. GMDR analysis suggested that the interaction model composed of the rs1800796 and obesity was the best model with statistical significance (P value from sign test [P] = 0.012), indicating a potential gene-environment interaction between rs1800796 and obesity. Overall, the two-locus models had a cross-validation consistency of 10/10 and had the testing accuracy of 0.641. We also conducted stratified analysis for rs1800796 genotype and obesity, and found that obese subjects with CG or GG genotype have the highest OST risk, compared to subjects with CC genotype, and normal BMI OR (95% CI) = 2.21 (1.52-3.49), after adjustment for age, smoke, and alcohol consumption status. CONCLUSIONS Our results suggested that the C allele of rs1800796 and the C allele of rs2069849 of IL-6 gene interaction between rs1800796 and abdominal obesity were all associated with increased OST risk.
Aged ; Aged, 80 and over ; China ; Female ; Gene-Environment Interaction ; Humans ; Interleukin-6 ; genetics ; metabolism ; Middle Aged ; Obesity ; epidemiology ; etiology ; genetics ; Osteoporosis ; epidemiology ; etiology ; genetics ; Polymorphism, Single Nucleotide ; Postmenopause ; genetics ; physiology ; Risk Factors

Objective: To assess clinical features and treatment outcomes in immunocompetent patients with primary central nervous system lymphoma (PCNSL). Methods: Sixty-two patients with PCNSL who attended Guangdong General Hospital between January 1998 and January 2012 were included. Survival curves were estimated using Kaplan-Meier survival methodology and statistical significance of continuous was assessed via the Cox proportional hazard model. Results: The median age of the patient cohort was 56 years, and the male to female ratio was 1.14∶1.00. The common presentations were increased intracranial pressure symptoms and neuron damage. Performance status of 54 (54/62, 87.1%) patients were the international prognostic index (IPI) 0-2. Diffuse large B-cell lymphoma (57/62, 91.9%) was most common, and the rest were T-cell lymphoma (4/62,6.4%) and extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (1/62, 1.6%). In the series, 32 patients (32/62, 51.6%) had multiple lesions. Involvement of deep structures was found in 30 (30/62, 48.4%) patients. An elevated serum LDH level was detected in 19 (19/62, 30.6%) patients and the Ki-67 index was ≥90% in 38 (38/62, 61.3%) patients. Univariate analysis showed patients who were female, age<60 years, had WHO Eastern Cooperative Oncology Group performance status grade 0-2, single lesion, absence of deep structures involvement and normal LDH level showed better 2-year survival rate and longer median survival time. Significance was only seen in the normal LDH level group. Multivariate Cox regression analysis revealed that radical surgery only and Rituximab+ high-dose of methotrexate+ whole brain radiation therapy (WBRT) were independent prognostic indicators in PCNSL patients (P<0.05). Conclusions PCNSL is a rare but aggressive tumor with poor prognosis. Patients treated with high-dose of methotrexate combining with rituximab, followed by WBRT have a better prognosis and longer survival time, and thus these could probably be a promising treatment.

assess clinical features and treatment outcomes in immunocompetent patients with primary central nervous system lymphoma (PCNSL). Methods Sixty?two patients with PCNSL who attended Guangdong General Hospital between January 1998 and January 2012 were included. Survival curves were estimated using Kaplan?Meier survival methodology and statistical significance of continuous was assessed via the Cox proportional hazard model. Results The median age of the patient cohort was 56 years, and the male to female ratio was 1.14∶1.00. The common presentations were increased intracranial pressure symptoms and neuron damage. Performance status of 54 (54/62, 87.1%) patients were the international prognostic index (IPI) 0-2. Diffuse large B?cell lymphoma (57/62, 91.9%) was most common, and the rest were T?cell lymphoma (4/62, 6.4%) and extranodal marginal zone lymphoma of mucosa?associated lymphoid tissue (1/62, 1.6%). In the series, 32 patients (32/62, 51.6%) had multiple lesions. Involvement of deep structures was found in 30 (30/62, 48.4%) patients. An elevated serum LDH level was detected in 19 (19/62, 30.6%) patients and the Ki?67 index was ≥90% in 38 (38/62, 61.3%) patients. Univariate analysis showed patients who were female, age<60 years, had WHO Eastern Cooperative Oncology Group performance status grade 0-2, single lesion, absence of deep structures involvement and normal LDH level showed better 2?year survival rate and longer median survival time. Significance was only seen in the normal LDH level group. Multivariate Cox regression analysis revealed that radical surgery only and Rituximab+ high?dose of methotrexate+ whole brain radiation therapy (WBRT) were independent prognostic indicators in PCNSL patients (P<0.05). Conclusions PCNSL is a rare but aggressive tumor with poor prognosis. Patients treated with high?dose of methotrexate combining with rituximab, followed by WBRT have a better prognosis and longer survival time, and thus these could probably be a promising treatment.

Objective: The objective of this study was to compare the early clinical effects of zirconia all⁃ceramic crowns using two different impression methods. Zirconia ceramic crowns were produced using digital models based on either a silicone rubber impression perfusion model in vitro or a 3D mouth scanner. Methods : A total of 50 patients with a planned restoration of the first permanent molar with zirconia all⁃ceramic crowns after root canal therapy were se⁃ lected and randomly divided into two groups: a digital impression by intraoral 3D scanning group and a digital impres⁃ sion by extraoral scanning after silicone rubber impression group. Zirconia all⁃ceramic crowns were created by CAD/ CAM in both groups. Marginal adaptation, proximal contact, and occlusal contact were compared between groups. Results: There was no significant difference between the two groups in marginal adaptation (P > 0.05). For proximal con⁃ tact and occlusal contact, no significant differences regarding the number of cases for the criteria of level A and level B were found between two groups (P > 0.05), while the misfit value in level B was smaller in the first group than in the second group (P < 0.05). Conclusion Zirconia all⁃ceramic crowns with intraoral 3D scanning show excellent early clinical performance.

Objective To examine the effects and mechanisms of siRNA targeting aquaporin 4 (AQP 4) in combination with hyperbaric oxygen therapy(HBO) on cerebral edema and apoptosis in the brain tissue of rats after hemorrhage.Methods Rats were randomly divided into four groups,the control group,the hyperbaric oxygen group,the AQP-4 siRNA group and the combination therapy group (24 rats).Thrombin Ⅶ was injected into the caudate nucleus to establish the hemorrhage model.Construction of siRNA targeting aquaporin 4 was conducted.The mRNA expression of AQP-4 was detected by RT-PCR at day 3.Changes in brain moisture and blood-brain barrier perme ability were measured by a wet/dry weight method and Evans blue fluorometry.The nerve cell apoptosis rate was analyzed by Annexin V andTdT-mediated dUTP-biotin nick end labeling (TUNEL).The expression of proteins including AQP-4,MMP-2,MMP-9,Bcl-2 and caspase-3 was detected by Western Blotting.All the animals were given a score for their nerve function at day 3.Results AQP-4 siRNA treatment obtained better effects than HBO in decreasing the brain edema leveland silencing AQP-4 mRNA(P＜0.05)while,the combination therapy group achieved the best results(P＜ 0.05).Compared with the control group,the percentage of apoptotic cells decreased in all the three treatment groups,with the most marked decrease observed in the combination treatment group(4.24± 0.04)％(F=13.76,P=0.001).The expression of AQP-4,MMP-2,MMP-9 and caspase-3 was lower (P＜0.05) and the expression of Bcl-2 was higher(P＜0.01)in the combination treatment group than in the other three groups.Compared with the control group,all the other three groups received better scores on nerve function defect evaluation at day 3 after hemorrhage(P＜0.05),with the combination treatment group again achieving the most favorable score (4.7 ± 1.1) (F=7.21,P =0.013).Conclusions Targeted siRNA interference combined with hyperbaric oxygen can effectively reduce cerebral edema after cerebral hemorrhage,inhibit neuronal apoptosis and promote neuron function recovery.The underlying mechanisms may be related to down-regulation of AQP-4,MMP 2,MMP-9 and caspase-3 expression and up-regulation of Bcl-2 expression.