[Advances in molecular mechanism and treatment of chronic mucus hypersecretion].

Chronic obstructive pulmonary disease (COPD) is one of the chronic diseases with high morbidity and mortality in China, which imposes heavy economic burden on society. Research has shown that chronic mucus hypersecretion (CMH) is an independent risk factor for persistent clinical symptoms, poor quality of life, rapid decline in lung function, acute exacerbation and increased hospitalization rate in COPD patients. CMH is a clinical phenotype of COPD with specific pathological and physiological changes. At present, the formation mechanism of CMH is not clear. There is a lack of specific and effective targeted treatments. This article aimed to review the latest research findings on CMH at home and abroad from the overview, impact on COPD patients, molecular mechanisms of formation, current treatment status and progress, and discuss potential targets for CMH treatment, to provide new ideas and directions for improving CMH and treating COPD.

[Characteristics and microRNA expression profile of exosomes derived from odontogenic dental pulp stem cells].

OBJECTIVE: To investigate the characteristics of exosomes derived from dental pulp stem cells (DPSCs) in the direction of odontogenic differentiation, to analyze the differences in microRNA expression profile between exosomes derived from undifferentiated and odontogenic DPSCs, and to analyze their possible signal transduction pathways. METHODS: (1) DPSCs were cultured in α minimum Eagle' s medium (α-MEM), and odontogenic DPSCs were cultured in odontogenic differentiation medium for 21 days, using alizarin red staining and alkaline phosphatase staining to identify the odontogenic differentiation. Exosomes from the cell supernatant were isolated respectively, named as dental pulp stem cells-exosomes (DPSCs-Exo) and dental pulp stem cells-odontogenic-exosomes (DPSCs-OD-Exo). The exosomes were identified by transmission electron microscopy, nanoparticle tracking analysis and Western blot. (2) The microRNA expression profiles of DPSCs-Exo and DPSCs-OD-Exo were investigated by microRNA microarray. To validate the result of the microRNA microarray, real-time quantitative polymerase chain reaction (real-time PCR) assay was applied on 3 most significantly differential expressed microRNA. Pathway analysis was taken to detect enriched pathways associated with the predicted target genes of microRNA. RESULTS: (1) The DPSCs were isolated and cultured in vitro showed typical fibroblast-like morphology. The odontogenic differentiated DPSCs were spindle-shaped, polygonal, and uniform in size. Odontogenic differentiation group showed a large number of dark deposits in alizarin red staining and the cells were darkly stained in alkaline phosphatase staining, while the cells in normal culture medium group did not show obvious dyeing. The DPSCs-Exo and DPSCs-OD-Exo had the same morphology, both showed bilayer membrane and cup-shape. The peak sizes of DPSCs-Exo and DPSCs-OD-Exo were (114.67±9.07) nm and (134.00±8.54) nm, respectively. The difference between the two was statistically significant. DPSCs-Exo and DPSCs-OD-Exo both expressed the markers of exosomes, tumor susceptibility gene (TSG)101 and CD63. (2) microRNA microarray results showed that the expression profiles of DPSCs-Exo and DPSCs-OD-Exo were different. Nineteen increased by more than two times, and one decreased by 64%. Real-time PCR results showed that the expression levels of microRNA-1246, microRNA-1246-100-5p and microRNA-1246-494-3p in DPSCs-OD-Exo were significantly up-regulated. The difference was statistically significant. microRNA target prediction database and gene signaling pathway database were used to analyze differentially expressed microRNA, and it was predicted that differentially expressed microRNA could target axis inhibition protein 2(AXIN2) gene and Wnt/ß-catenin signaling pathway. CONCLUSION: DPSCs-OD-Exo and DPSCs-Exo had differences in their microRNA expression profile. Those differentially expressed microRNA may be involved in the regulation of DPSCs odontogenic differentiation.

[Oligonucleotide drugs and their progress in stomatology].

Oligonucleotide drugs have the characteristics of targeting, modifiability and high biosafety. Recent studies have shown that oligonucleotide can be used to make biosensors, vaccine adjuvants, and has the functions of inhibiting alveolar bone resorption, promoting jaw and alveolar bone regeneration, anti-tumor, destroying plaque biofilm, and precise control of drug release. Therefore, it has a broad application prospect in the field of stomatology. This article reviews the classification, action mechanism and research status of oligonucleotide in stomatology. The aim is to provide ideas for further research and application of oligonucleotide.

[Oligonucleotide drugs and their progress in stomatology].

Oligonucleotide drugs have the characteristics of targeting, modifiability and high biosafety. Recent studies have shown that oligonucleotide can be used to make biosensors, vaccine adjuvants, and has the functions of inhibiting alveolar bone resorption, promoting jaw and alveolar bone regeneration, anti-tumor, destroying plaque biofilm, and precise control of drug release. Therefore, it has a broad application prospect in the field of stomatology. This article reviews the classification, action mechanism and research status of oligonucleotide in stomatology. The aim is to provide ideas for further research and application of oligonucleotide.

[Pathogenesis and classification of tooth resorption].

Tooth resorption is an idiopathic destructive disease of dental hard tissues. The etiology and pathogenesis remain obscure. It has various manifestations and can be commonly classified as internal tooth resorption and external root resorption on the basis of the resorptive lesion sites. There have been many attempts to make further classification based upon the pathological manifestations in recent years. Radiographic examination is an effective tool to assist in the diagnosis. There are few systematic researches on tooth resorption worldwide, most of which are case reports. This review elaborates on the research progress of tooth resorption from aspects of pathogenesis and classification.

[Association between weight gain during the first half of pregnancy and the risk of hypertension disorder of pregnancy: a prospective cohort study].

Objective: To explore the association between weight gain during the first half of pregnancy and the risk of hypertension disorder of pregnancy (HDP). Methods: This prospective cohort study recruited singleton pregnant women in the first trimester from November 2016 to March 2019 at 19 community hospitals in Tianjin. According to pre-pregnancy body mass index (BMI), the cohort was divided into 3 groups: underweight(BMI<18.5 kg/m2), normal-weight(18.5-24.9 kg/m2), and overweight/obese(≥25.0 kg/m2). The basic information of the participants was gathered through questionnaires, and the height, weight, and blood pressure of the participants were measured along with routine pregnancy examinations. The rate of gestational weight gain (rGWG) in the 3 periods (0-13+6, 14+0-20+6, and 0-20+6 weeks) of the participants was calculated. To observe the occurrence of HDP, the participants were followed up to 42 days postpartum. Using a generalized linear model, the association between rGWG at the 3 periods during the first half of pregnancy and HDP after 20 weeks of gestation was evaluated. Results: A total of 9 805 pregnant women were finally included, with the age of (30.6±3.8) years old, 9 418 (96.1%) Han ethnicity, and 6 845 (69.8%) primipara. There were 1 184 (12.1%), 6 831 (69.7%) and 1 790 (18.3%) participants in the underweight, normal-weight, and overweight/obese groups. Five hundreds and eight pregnant women were diagnosed with HDP (5.2%). The incidences of HDP were 1.8% (21/1 184), 3.9% (269/6 831), and 12.2% (218/1 790), respectively, in underweight, normal-weight, and overweight/obese groups. Adjusted for age, pre-pregnancy BMI, primipara, and family history of hypertension, women in the entire cohort with rGWG ≥ 0.18 kg/week before 13+6 weeks of pregnancy had a 28% higher HDP risk than those with rGWG ≤ 0.00 kg/week (RR=1.28, 95%CI 1.04-1.55, P=0.015), and the risk of HDP was increased by 39% in the overweight/obese group (RR=1.39, 95%CI 1.04-1.85, P=0.026), while no correlation was found between rGWG and HDP in underweight and normal-weight pregnant women (P>0.05). Weight gain during 14+0-20+6 weeks of pregnancy in any group was not related to the risk of HDP (P>0.05).In the entire cohort, compared to rGWG ≤0.14 kg/week, rGWG≥0.28 kg/week prior to 20+6 weeks increased HDP risk by 36% (RR=1.36, 95%CI 1.11-1.67, P=0.003). Normal-weight pregnant women with rGWG≥0.29 kg/week faced a 46% higher risk of HDP than those with rGWG≤0.15 kg/week (RR=1.46, 95%CI 1.11-1.93, P=0.008).In the overweight/obese group, excessive weight gain before 20+6 weeks seemed to increased risk of HDP, but the difference was not statistically significant (RR=1.35,95%CI 0.99-1.85, P=0.059), while the connection was nonexistent in underweight women. Conclusions: Except for pre-pregnancy underweight women, excessive weight gain during the first half of pregnancy is associated with increased risk of HDP among pregnant women.

Osteocalcin reduces fat accumulation and inflammatory reaction by inhibiting ROS-JNK signal pathway in chicken embryonic hepatocytes.

Osteocalcin (OCN) has a function in preventing fatty liver hemorrhagic syndrome (FLHS) in poultry. The aim of this study was to investigate the effects of OCN on fat emulsion stimulated chicken embryonic hepatocytes and related signaling pathways. The primary chicken embryonic hepatocytes were isolated from the incubated 15-day (E15) pathogen free eggs and cultured with dulbecco's modified eagle medium (DMEM). After the hepatocyte density reached 80%, the cells were divided into 5 groups: control group (CONT), fat emulsion group (FE, 10% FE, v/v), FE with ucOCN at 1 ng/mL (FE-LOCN), 3 ng/mL (FE-MOCN), and 9 ng/mL (FE-HOCN). In addition, 2 mM N-Acetyl-L-cysteine (NAC) a reactive oxygen species (ROS) scavenger, and 5 µM SP600125, a Jun N-terminal kinase (JNK) inhibitor, were added separately in to the DMEM with 10% FE to test effects of FE on the function of ROS-JNK signal pathway. The number of hepatocytes, cell ultra-microstructure, viability, and apoptosis were detected after 48 h treatment, and the protein expressions and enzyme concentrations were detected after 72 h treatment. The results showed that, compared to the control group, FE increased the triglyceride (TG) concentration and lipid droplets (LDs) in chicken embryonic hepatocytes (P < 0.05), and induced hepatocytic edema with obviously mitochondrial swelling, membrane damage, and cristae rupture. FE also decreased ATP concentration, increased ROS concentrations and mitochondrial DNA (mtDNA) copy number, promoted inflammatory interleukin-1 (IL-1), IL-6, tumor necrosis factor-alpha (TNF-α) concentrations and hepatocytic apoptosis rate, and raised phospho-c-Jun N-terminal kinase (p-JNK) protein expressions. Compared to the FE group, ucOCN significantly increased hepatocyte viability, reduced hepatocytic TG concentrations and LDs numbers, and alleviated hepatocytic edema and mitochondrial swelling. Furthermore, ucOCN significantly decreased ROS concentrations, increased ATP concentrations, reduced IL-1, IL-6, TNF-α concentrations and hepatocytic apoptosis rate, and inhibited p-JNK protein expressions (P < 0.05). NAC had the similar functions of ucOCN reduced the ROS concentration and inhibited the TNF-α protein expression and p-JNK/JNK ration. Similarly, SP600125 reduced p-JNK/JNK protein expression, IL-1, IL-6, TNF-α, and TG concentrations without effects on ROS concentration and hepatocytic apoptosis. These results suggest that ucOCN alleviates FE-induced mitochondrial damage, cellular edema, and apoptosis of hepatocytes. These results reveal that the functions of ucOCN in reducing fat accumulation and inflammatory reaction in chicken embryonic hepatocytes are mostly via inhibiting the ROS-JNK signal pathway.

Bacillus subtilis inhibits intestinal inflammation and oxidative stress by regulating gut flora and related metabolites in laying hens.

Bacillus subtilis is one of the most popular commercial probiotics used in farm animal production. However, its potential mechanisms are not very clear. The aim of this study was to investigate the effects of dietary Bacillus subtilis on intestinal histomorphology, innate immunity, microbiota composition, transcriptomics, and related metabolomics. Twenty-four 48-week-old Lohman Pink-shell laying hens were randomly divided into two groups: a basic diet and the basic diet supplemented with Bacillus subtilis (0.5 g/kg) for a 9-week experiment. At the end of the experiment, tissues of the duodenum, ileum, and jejunum as well as cecal content of each bird were collected for microstructure, PCR, transcriptome, metabolome, and 16S rRNA analyses. The results showed that dietary Bacillus subtilis supplement had no effect on the intestinal microstructure. However, Bacillus subtilis increased mRNA expression of tight junction protein occludin (P < 0.05), while reduced mRNA expression of lipopolysaccharide-induced TNF factor (P < 0.01) in the duodenum. Moreover, transcriptomic results indicated that most of Bacillus subtilis supplement-induced differential genes were associated with inflammation and immunity, including cytochrome b-245 beta chain, transferrin, and purinergic receptor P2X 7, resulting in a decrease in Malondialdehyde level (P < 0.05) in the duodenum. In addition, at the genus level, Bacillus subtilis supplement enriched the potential beneficial bacteria, Candidatus_Soleaferrea (P = 0.02) but inhibited the harmful bacteria including Lachnospiraceae_FCS020_group, Ruminiclostridium, Lachnospiraceae_UCG-010, and Oxalobacter. Metabolomic results revealed that N-Acetylneuraminic acid and ADP were increased by fed Bacillus subtilis. These results suggest that dietary Bacillus subtilis could inhibit gut inflammation and improve antioxidative status and barrier integrity of the duodenum via regulating gut microbial composition in laying hens.

[Biological basis and clinical exploration of regenerative endodontic therapy].

Regenerative endodontic therapy is a tissue engineering based approach of treatment for endodontic disease. Its purpose is to achieve the regeneration of the pulp-dentin complex, thus to promote root development of the immature permanent tooth with necrotic pulp. Like other treatments based on tissue engineering techniques, the success of regenerative pulp therapy depends on such three elements as seed cells, scaffold materials and growth factors. Since its inception 20 years ago, there have been various terminologies in the literature, with similarities and differences in connotation. The present article summarizes and analyzes the term evolution, biological basis, clinical considerations and future scientific research directions of regenerative endodontics, in order to find out the unsolved scientific problems and to promote the development and standardization of this technique in clinical practice.

[Research progress in the application of dopa-inspired compounds to improve the dentin-resin bonding].

Mussle foot protein has a main component which is named dopa. Dopa can be used to promote a relatively firm adhesion of mussels to the surface of solid materials through forming dihydrogen bonds, π-π/π-cation bonds and chelating metals,etc. To exploit these interactions, there is the opportunity to apply dopa-inspired compounds to improve the dentin-resin bonding. The current review provides valuable information concerning the mechanism of adhesion mediated by mussel foot protein and describes the application of dopa-inspired compounds in the dentin-resin bonding. The article provides novel information for future research in optimization of the properties of dentin-resin bonding.

Osteocalcin prevents insulin resistance, hepatic inflammation, and activates autophagy associated with high-fat diet-induced fatty liver hemorrhagic syndrome in aged laying hens.

The aim of this study was to investigate the effects of osteocalcin (OCN) on fatty liver hemorrhagic syndrome (FLHS) in aged laying hens. Thirty 68-week-old White Plymouth laying hens were randomly assigned into conventional single-bird cages, and the cages were randomly allocated into one of 3 treatments (n = 10): normal diet (ND + vehicle, ND + V), high-fat diet (HFD + vehicle, HFD + V), and HFD + OCN (3 µg/bird, 1 time/2 d, i.m.) for 40 d. At day 30, oral glucose tolerance tests (OGTT) and insulin tolerance tests (ITT) were performed. At the end of experiment, the hens were euthanized followed by blood collection. The plasma aspartate transaminase (AST), alkaline phosphatase (ALP), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were measured using an automatic biochemistry analyzer. Pathological changes in the liver were examined under both light and transmission electron microscopes. The plasma inflammatory factors including interleukin-1 (IL-1), IL-6, and tumor necrosis factor-alpha (TNF-α) were analyzed by ELISA, and the gene expressions of these inflammatory factors in the liver were analyzed by real-time PCR. The level of oxidative stress was evaluated using malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) assay kits, respectively. The results showed that HFD + V hens had more severe liver hemorrhage and fibrosis than ND + V hens (P < 0.05). The ultramicrostructural examination showed that hepatocytes of HFD + V hens exhibited necrotic pyknosis showing great intracellular electron, mitochondrial swelling, shrunk nucleus, and absence of autolysosomes. Osteocalcin mitigated HFD + V-induced pathological changes in aged laying hens. High-fat diet + OCN hens had higher insulin sensitivity; lower liver concentrations of MDA (P = 0.12) but higher GSH-Px (P < 0.05); and lower blood TNF-α concentrations (P < 0.05) and mRNA expressions (P < 0.05) than HFD + V hens. These results suggest OCN functions in preventing the FLHS process in old laying hens through inhibiting excessive energy diet-induced metabolic disorder, oxidative stress, and related pathological damage.

[Application of penile index in the diagnosis and therapeutic evaluation of concealed penis in children].

Objective: To explore the significance of penile index in the diagnosis of concealed penis and the evaluation of therapeutic efficacy. Methods: A retrospective study was conducted on 221 children with phimosis and 113 children with concealed penis aged 6-10, all of whom had undergone circumcision or phalloplasty respectively in Shanghai Children's Medical Center from January 2014 to December 2017. Penile index was measured before and after surgery. The values of phimosis and concealed penis were tested by group t test. Self -control test was performed for values of concealed penis before and after operations. Results: Penile index was 0.78±0.08 in children with phimosis. It was 0.23±0.10 in concealed penis before operation and changed to 0.84±0.11 postoperatively. In children with phimosis and concealed penile, the difference of penile index was statistically significant (P<0.001). The difference of penile index before and after operation was statistically significant (P<0.001) in children with concealed penis. Conclusion: Penile index is an effective index to evaluate the degree of penis exposure.

[Mediated pathways of exosomes uptake by stem cells of apical papilla].

OBJECTIVE: To evaluate the uptake of exosomes by stem cells from apical papilla (SCAP), thus to provide experimental basis for mechanism of the exosomes endocytosis by SCAP. METHODS: (1) Exosomes of dental pulp stem cells (DPSCs) were isolated by hypercentrifugation combined with ultrafiltration method. The exosomes were identified by transmission electron microscopy, nanoparticle tracking analysis and western blot. (2) PKH-26 membrane labeling technology was used to mark the DPSCs derived exosomes. The labeled exosomes were co-cultured with SCAP at 37 °C as positive control group, and co-cultured with SCAP at 4 °C as the low-temperature treatment group, while the negative control group was set up. (3) Using clathrin-mediated endocytosis inhibitor chlorpromazine (CPZ, 10 µmol /L) as CPZ group, caveolae-mediated endocytosis Genistein (200 µmol/L) as Genistein group, and macropinocytosis inhibitor LY294002 (50 µmol/L) as LY294002 group to treat the SCAP respectively. Solvent control group (DMSO group) was set. Immunofluorescence staining was used to detect the red fluorescence SCAP and flow cytometry was used to analyze the proportion of SCAP labeled with red fluorescence. RESULTS: (1) The bilayer membrane and cup-shaped appearance of representative exosomes were observed. The peak of the size of DPSCs-derived exosomes was at 144 nm. The exosomes expressed exosomal marker proteins TSG101 and CD63, but not GAPDH which was the cellular internal control protein. (2) Immunofluorescence staining showed that after being co-cultured at 37 °C for 6 hours, red fluorescence could be detected in SCAP but it could not be detected after being co-cultured at 4 °C for 6 hours. After endocytosis inhibition, the red fluorescence in SCAP was reduced. Flow cytometry showed that the proportion of SCAP labeled with red fluorescence in positive group was 35.0%, in negative control group was 0.5%, and in solvent control group was 29.7%, in CPZ group, Genistein group and Genistein group were reduced to 13.7%, 16.6%, and 20.9%, respectively. CONCLUSION: SCAP could uptake the DPSCs derived exosomes, and low temperature could inhibit this process. The exosomes uptake of SCAP was mediated by the clathrin endocytosis pathway, caveolae-mediated endocytosis and macropinocytosis pathway.

Expression of Normal or Mutated X-Linked BCOR Transcripts in OFCD iPSCs.

Reprogramming diseased cells with mutated genes into induced pluripotent stem cells (iPSCs) can allow studies of disease mechanism and correct the mutation. Oculofaciocardiodental (OFCD) syndrome is a developmental disorder caused by heterozygous mutations in the X-linked BCL-6 corepressor (BCOR) gene. In this present study, we aimed to reprogram stem cells from a tooth apical papilla (SCAP) of a patient with OFCD, termed SCAP-O, into iPSCs. The SCAP-O carry a copy of the BCOR gene having 1 nucleotide deletion in 1 of the alleles, therefore harboring a mixture of cells expressing either normal (SCAP-OBCOR-WT) or mutated (SCAP-OBCOR-mut) BCOR transcripts. We subcloned SCAP-O and separated SCAP-OBCOR-WT and SCAP-OBCOR-mut as verified by sequencing. The selected subclone SCAP-OBCOR-mut expressed only the mutated BCOR transcripts and remained in such condition after multiple passages. We reprogrammed SCAP-O and subclone SCAP-OBCOR-mut into transgene-free iPSCs using an excisable lentiviral vector system (hSTEMCCA-loxP) carrying 4 reprogramming factors in a single cassette, followed by removal of transgenes via Cre-mediated excision. We found that after reprogramming SCAP-O or subclone SCAP-OBCOR-mut into iPSCs, some of the iPSC clones expressed either solely the normal BCOR-WT or BCOR-mut transcripts, while other clones expressed both BCOR-WT and BCOR-mut transcripts. This is our first step toward establishing OFCD study models by generating isogenic control BCOR-WT iPSCs versus BCOR-mut iPSCs.

[Role of endocytosis in cell surface CXC chemokine receptor 4 expression of stem cells from apical papilla].

OBJECTIVE: To evaluate the change of cell surface CXC chemokine receptor 4 (CXCR4) expression of stem cells from apical papilla (SCAP) after the inhibition of endocytotic pathway, thus to provide experimental basis for the mechanism of SCAP migration. METHODS: The immunofluorescence analysis was conducted to examine the co-expression of CXCR4 and endocytotic compartments, including early endosomes, recycling endosomes and lysosomes in SCAP. Several Rab proteins were applied as markers of organelles in the endocytotic pathway, including Rab5 for early endosomes, Rab11A for recycling endosomes, and Lamp1 for lysosomes. The co-localization of CXCR4 with these endodontic compartments was further observed by proximity ligation assay (PLA). SCAP was treated with two kinds of endocytotic inhibitors, Blebbistatin and Dynasore, at a concentration of 80 µmol/L, respectively. The conditioning time was 1 hour. Flow cytometry was carried out to evaluate the proportion of SCAP that expressed CXCR4 on cell surface. The data were analysed by analysis of variance (ANOVA). RESULTS: The red staining of CXCR4 on immunofluorescence confocal microscopy predominantly overlapped with the green staining of Rab5 and Rab11A, and partly overlapped with Lamp1. It indicated that most CXCR4 molecules were located in early endosomes and recycling endosomes, and some were located in lysosomes. The PLA results revealed that the co-localizaiton of CXCR4 with endocytotic compartments could be observed in early endosomes, recycling endosomes and lysosomes. According to the results of flow cytometry, the proportion of SCAP that expressed CXCR4 on cell surface was as low as 0.13%±0.10%. After the inhibition of endocytosis by pretreating the cells with the following two inhibitors, Blebbistatin and Dynasore, the percentage of SCAP that positively expressed CXCR4 on cell surface was significantly increased to 13.34%±1.31% in Blebbistatin group and 4.03%±0.92% in Dynasore group (F=16.721, P<0.001). Moreover, the number of SCAP that expressed CXCR4 on cell surface in Blebbistatin group was significantly higher than that in Dynasore group (P<0.001). CONCLUSION: The inhibition of endocytotic pathway could increase the number of SCAP that expressed CXCR4 on cell surface, and provide potency for the migration of SCAP.

[Difference of one year death and stroke recurrence in ischemic stroke patients with anterior and posterior circulation intracranial atherosclerosis].

Objective: To explore the differences of one year death and stroke recurrence between ischemic stroke patients with intracranial atherosclerotic stenosis or occlusion of anterior circulation and those of posterior circulation. Methods: All the patients were from the Chinese Intracranial Atherosclerosis Study (CICAS), between October 2007 and June 2009; patients with extracranial stenosis or occlusion, patients without acute infarction by diffusion weighted image, and patients with intracranial atherosclerosis of both anterior and posterior circulation were excluded.All the enrolled patients were divided into three groups: no significant intracranial atherosclerosis group (n=964), anterior circulation intracranial atherosclerosis group (n=440), posterior circulation intracranial atherosclerosis group (n=233). One year outcome was evaluated by any cause of death and stroke recurrence. Results: Of the 1 637 patients, 30 cases were died and 58 cases had stroke recurrence within one year.Compared with : no significant intracranial atherosclerosis group, adjusted hazard ratio (95% confidence interval) of one-year death for anterior and posterior circulation intracranial atherosclerosis group were 1.349 (0.311-5.851), 4.542 (1.227-16.813), respectively.Adjusted hazard ratio (95% confidence interval) of one year stroke recurrence were 1.663 (0.620-4.460) and 2.464 (0.935-6.493), respectively. Conclusions: Ischemic stroke patients with intracranial atherosclerosis of posterior circulation has higher risk of one year death. One year stroke recurrence risk for patients with intracranial atherosclerosis of anterior and posterior circulation needs to be further evaluated.

Effects of ALA-PDT on HPV16-immortalized cervical epithelial cell.

Current treatments for high-grade cervical intraepithelial neoplasia (CIN) and persistent infection with high-risk human papillomavirus (HR-HPV) type are mainly surgical interventions. However, such treatments are associated with adverse side effects and pose risks for future pregnancies. In order to reduce the requirement for excisional procedures, an effective and noninvasive therapy is needed for women at reproductive age. ALA-PDT has proved to be effective in the treatment of HPV-associated disease in several clinical investigations. In this study, the anti-proliferative effect of ALA-PDT was investigated in HPV16-immortalized cervical epithelial H8 cells. CCK-8 assay was used to measure cytotoxicity in H8 cells. The IC50 of ALA-PDT on H8 cells was about 120.75 ± 1.18 µM. We have now evaluated the mechanism by which ALA-PDT induces cell death. Annexin V-FITC/PI staining showed a significant dose-dependent induction of apoptosis by ALA-PDT in H8 cells, associated with accumulation of the tumor suppressor protein p53 and the cyclin-dependent kinase inhibitor p21. Furthermore, ALA-PDT down-regulates expression of HPV E6/E7 oncogene as well as up-regulate tumor suppressor RbAp48 protein. Together, our data provides a basis for understanding and developing ALA-PDT as a cure for HPV infection-associated diseases and prevention of cervical cancer.

[Effect of root canal sealers on biocompatibility of human periodontal ligament cells].

OBJECTIVE: To compare the effects of three root canal sealers with respect to time on biocompatibility of human periodontal ligament cells (hPDLCs).The sealers included zinc oxide and eugenol based sealers (ZOE), epoxy resin sealers (ERS) and silicone based sealers (SBS). METHODS: hPDLCs were primarily cultured,with the method combining of tissue explant and enzymatic digestion. The cells were then exposed to different extract fluids: (1) ZOE extracted for 24 h group ;(2) ZOE extracted for 1 week group;(3)ZOE extracted for 2 weeks group;(4) ERS extracted after 24 h group; (5) ERS extracted after 1 week group; (6) ERS extracted after 2 weeks group; (7) SBS extracted after 24 h group; (8) SBS extracted after 1 week group; (9) SBS extracted after 2 weeks group; (10) Dulbecco modified Eagle's medium/F12 (DMEM/F12) as negative control group. Cell morphology was observed under an inverted microscope.Cell proliferation was measured by methyl-thiazol-diphenyltetrazolium (MTT) assay. ALP assay kit was used for measuring alkaline phosphatase (ALP) activity. Sealers of 2 weeks' setting time were then immersed in an osteogenic medium for examination of mineral nodules and calcium deposits. RESULTS: Considering the relative growth rate(RGR),ZOE was severely to moderately cytotoxic(RGR:13.6%-39.9%), while ERS was slightly or not cytotoxic (RGR: 87.6%-95.3%).Only SBS did not show any cytotoxicity after setting (RGR: 91.8%-106.7%). The setting time influenced the cytotoxicity of ERS which decreased after 1 week. Considering the ALP activity,there was no difference between SBS group and control group (F=3.397,P=0.053). According to the results of calcium deposits, ZOE:D562 nm=0.180±0.050,ERS: D562 nm=2.968±0.201,SBS:D562 nm=3.623±0.039,Control:D562 nm=3.477±0.102,the ranking of ALP activity and calcium deposits was as follows: ZOE

Clinical significance of sCIP2A levels in breast cancer.

OBJECTIVE: It has previously found that human oncoprotein cancerous inhibitor of protein phosphatase 2A (CIP2A) was overexpressed in breast cancer, and was positively correlated with lymph node metastasis of the patients. This study aimed to investigate the association between serum CIP2A and prognosis of breast cancer. Then, we investigated whether CIP2A could be as a therapeutic target in breast cancer treatment. PATIENTS AND METHODS: Preoperative CIP2A levels of 240 patients with breast cancer and 480 cases of controls were measured by ELISA method. The association of CIP2A levels with clinicopathological outcomes was investigated by univariate and multivariate analyses. The effect of CIP2A on breast cancer MDA-MB-231 cells was evaluated by CIP2A siRNA-mediated depletion of the CIP2A protein followed by an analysis of cell proliferation, invasion, colony growth, and xenograft growth and metastasis. RESULTS: The serum CIP2A levels in patients with breast cancer were (79.0 ± 74.2) ng/mL, which was significantly higher than that in those controls (25.6 ± 21.4) ng/mL for male and (24.8 ± 20.6) ng/mL for female control. Higher preoperative CIP2A levels were significantly associated with the American Joint Committee on Cancer (AJCC) stage, histological grade and lymph node metastasis. Patients with elevated CIP2A levels showed worse survival. In multivariate analysis, elevated preoperative CIP2A levels were independent prognostic factors. Patients with high CIP2A levels had significantly shorter overall survival (OS) and disease-free survival (DFS) times. Knockdown of CIP2A by stable CIP2A siRNA transfection inhibited MDA-MB-231 cell proliferation, invasion, colony growth in vitro, and xenograft growth and metastasis in vivo. CONCLUSIONS: Our results suggest that serum CIP2A is significantly higher in patients with breast cancer, which is a potential biomarker to make a distinction between breast cancer patients and healthy controls. Higher serum CIP2A levels positively associated with the aggressive phenotype of breast cancer, and forecasts poor prognosis for patients with breast cancer. Knockdown of CIP2A may be a novel target for prevention and treatment of breast cancer.

Association of the glucocerebrosidase N370S allele with Parkinson's disease in two separate Chinese Han populations of mainland China.

BACKGROUND AND PURPOSE: Mutations in the glucocerebrosidase (GBA) gene have been implicated in the development of Parkinson's disease (PD). However, recent screenings for GBA mutations in PD subjects from different ethnic populations have yielded contradictory results. METHODS: We performed a case-control study to look for a possible association between PD and the GBA N370S allele involving 628 subjects in two separate Chinese Han populations from mainland China. All subjects were successfully genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: A total of six patients with PD and two control subjects carried the N370S allele. Although PD cases (1.8%) had an increased frequency of N370S compared to controls (0.7%), the difference was not statistically significant (P = 0.290). However, when PD cases were stratified by age at onset, a higher frequency of N370S in late-onset PD (LOPD) cases (3.2%) compared to controls was observed. CONCLUSIONS: Our results suggest that the N370S allele might be associated with LOPD in Chinese Han population and that this phenomenon should be further examined in a larger study.