Objective To explore the role of the PI3K/Akt signaling pathway in the calvarial osteolysis induced by TCP wear particles in mice model.Methods Thirty-six male ICR mice were randomly divided into a sham group (n=12),TCP group (n=12)and a LY294002-treated group (n=12).A murine calvarial model of osteolysis was established through implanting 30 mg of TCP particles onto the surface of bilateral parietal bones following the removal of the periosteum.On the second postoperative day,LY294002 (5 mg·kg-1)was locally injected to the calvarium under the periosteum three times a week;mice in the sham group received local injection of normal saline (N.S.)in the calvarium,and the injection time was consistent with that of LY294002.Two weeks later,the calvaria and periostea were obtained after the mice were executed.The calvarial osteolysis,bone mineral density (BMD)and bone mineral content(BMC)were analyzed using Micro-CT,Hematoxylin-Eosin (HE)staining was conducted to observe the inflammatrory response and formation of osteoclasts.Real-time PCR was applied to detect the mRNA level of tartrate-resistant acid phosphatase (TRAP),the marker of osteoclasts formation,cathepsin K (CstK),receptor activator for nuclear factor-κB kigand (RANKL)and c-Fos.The release of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6)and IL-1β were measured using enzyme-linked immumsorbent assay (ELISA).Results Micro-CT and histological analysis indicated that LY294002,the specific inhibitor of PI3K,significantly prevented TCP wear particles-induced osteolysis and osteoclastogenesis,and increased BMD and BMC in the calvaria of mice.Real-time PCR data revealed LY294002 significantly suppressed the increase in mRNA level of osteoclastogenic genes such as TRAP,CstK,RANKL and c-Fos in the calvaria of TCP wear particles-implanted group.ELISA assay showed that TCP wear particles-induced release of TNF-α,IL-1β and IL-6 was significantly inhibited by LY294002 treatment.Furthermore,LY294002 significantly attenuated TCP wear particles-triggered activation of Akt,and down-regulated the level of p-AktSer473 and p-AktThr308.Conclusion PI3K/Akt signaling pathway contributes to TCP wear particle-induced osteolysis,and can be developed as a new therapeutic target for the prevention and treatment of bone destruction diseases caused by wear debris.

To obtain a primary overview of gene diversity and expression pattern in Lycoris longituba, 4,992 ESTs (Expressed Sequence Tags) from L. longituba bud were sequenced and 4,687 cleaned ESTs were used for gene expression analysis. Clustered by the PHRAP program, 967 contigs and 1,343 singlets were obtained. Blast search showed that 179 contigs and 227 singlets (totally 1,066 ESTs) had homologues in GenBank and 3,621 ESTs were novel.
Base Composition ; Computational Biology ; Expressed Sequence Tags ; Flowering Tops ; genetics ; Gene Expression Regulation, Plant ; Genetic Variation ; Lycoris ; genetics

OBJECTIVETo observe the toxicity of Pulsatilla chinensis (Bunge) Regel saponins (PRS) against Oncomelania hupensis (O. hupensis).

METHODSO. hupensis snails were exposed to 40% and 80% of 24 h LC50 of PRS for 24 h, and then choline esterase (CHE), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activities in cephalopodium and liver of snails were determined. Niclosamide (NIC) was used as the reference molluscicide. Zebra fish lethality test was evaluated to non-target aquatic species of PRS.

RESULTSThe molluscicidal activity of PRS (LC50 at 24 h: 0.48 mg/L) was similar to that of NIC (LC50 at 24 h: 0.16 mg/L). Significant alterations about CHE, ALP, and ALT activities both in the cephalopodium and the liver of snails were observed when O. hupensis was exposed to 40% and 80% LC50 of PRS or NIC for 24 h. PRS and NIC could not affect LDH activity in the cephalopodium and the liver. Lower toxicity to fish of PRS was observed up to the highest concentration tested than NIC.

CONCLUSIONPRS, as compared with the reference molluscicide NIC, is thought to be used for the control of harmful vector snails safely.

Animals ; Molluscacides ; pharmacology ; Pulsatilla ; chemistry ; Saponins ; pharmacology ; Snails ; drug effects

PURPOSE: In human subjects and animal models with acute and chronic lung injury, the bioactive lysophosphatidylcholine (LPC) is elevated in lung lining fluids. The increased LPC can promote an inflammatory microenvironment resulting in lung injury. Furthermore, pathological lung conditions are associated with upregulated phospholipase A2 (PLA2), the predominant enzyme producing LPC in tissues by hydrolysis of phosphatidylcholine. However, the lung cell populations responsible for increases of LPC have yet to be systematically characterized. The goal was to investigate the LPC generation by bronchial epithelial cells in response to pathological mediators and determine the major LPC species produced. METHODS: Primary human bronchial epithelial cells (NHBE) were challenged by vascular endothelial growth factor (VEGF) for 1 or 6 h, and condition medium and cells collected for quantification of predominant LPC species by high performance liquid chromatography-tandem mass spectrometry (LC-MS-MS). The cells were analyzed by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) for PLA2. The direct effects of LPC in inducing inflammatory activities on NHBE were assessed by transepithelial resistance as well as expression of interleukin-8 (IL-8) and matrix metalloproteinase-1 (MMP-1). RESULTS: VEGF stimulation of NHBE for 1 or 6 h, significantly increased concentrations of LPC16:0, LPC18:0, and LPC18:1 in condition medium compared to control. The sPLA2-selective inhibitor (oleyloxyethyl phosphorylcholine) inhibited the VEGF-induced release of LPC16:0 and LPC18:1 and PLA2 activity. In contrast, NHBE stimulated with TNF did not induce LPC release. VEGF did not increase mRNA of PLA2 subtypes sPLA2-X, sPLA2-XIIa, cPLA2-IVa, and iPLA2-VI. Exogenous LPC treatment increased expression of IL-8 and MMP-1, and reduced the transepithelial resistance in NHBE. CONCLUSIONS: Our findings indicate that VEGF-stimulated bronchial epithelial cells are a key source of extracellular LPCs, which can function as an autocrine mediator with potential to induce airway epithelial inflammatory injury.
Epithelial Cells* ; Group X Phospholipases A2 ; Humans ; Hydrolysis ; Interleukin-8 ; Lung ; Lung Injury ; Lysophosphatidylcholines* ; Mass Spectrometry ; Matrix Metalloproteinase 1 ; Models, Animal ; Phosphatidylcholines ; Phospholipases A2 ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger ; Vascular Endothelial Growth Factor A

PURPOSE: Asthma is a chronic inflammatory disease of the airways, and is associated with upregulation of phospholipase A2 (PLA2), the enzyme that hydrolyzes phosphatidylcholine, producing lysophosphatidylcholine (LPC) and free fatty acids. LPC is a lipid mediator with known pro-inflammatory and pro-atherogenic properties, and is believed to be a critical factor in cardiovascular diseases. We postulate that asthmatic subjects have an elevated content of LPC in the lung lining fluids. METHODS: Eight non-asthmatic controls and seven asthmatic subjects were recruited for broncho-alveolar lavage fluids (BALF) collection for analysis of LPC by high performance liquid chromatography-tandem mass spectrometry. RESULTS: LPC16:0 and LPC18:0 were significantly elevated in the BALF of asthmatics with impaired lung function characteristic of moderate asthma, but not mild asthma. The increased LPC content in BALF was accompanied by increased PLA2 activity. Furthermore, qRT-PCR analysis of the BALF cell fraction indicated increased secretory PLA2-X (sPLA2-X). CONCLUSIONS: The increased LPC content in the lung lining fluids is a potential critical lipid mediator in the initiation and/or progression of airway epithelial injury in asthma.
Asthma ; Cardiovascular Diseases ; Fatty Acids, Nonesterified ; Lung* ; Lysophosphatidylcholines* ; Mass Spectrometry ; Phosphatidylcholines ; Phospholipases A2 ; Therapeutic Irrigation ; Up-Regulation

OBJECTIVETo study the relationship between the efficacy of interferon-alpha and the variation of perforin protein expression in the peripheral blood mononuclear cells in 35 patients with chronic hepatitis B (CHB).

METHODSThe perforin protein in the peripheral blood mononuclear cells was detected by immunocytochemistry technique.

RESULTSThe level of the perforin protein expression in the peripheral blood mononuclear cells was significantly higher in the post-treatment group with interferon-alpha than in the pre-treatment group. At the end of the treatment with interferon-alpha, there were 12 cases of complete responders, 14 cases of partial responders, and 9 cases of non-responders. After interferon-alpha treatment, the mean level of the perforin protein expression in the peripheral blood mononuclear cells was 12.1%, 6.9% and 3.9% respectively, and there existed significant differences among the three groups. Moreover, before treatment, the level of the perforin protein expression in the complete responder group was significantly higher compared to the partial responder group or the non-responder group.

CONCLUSIONTreatment with interferon-alpha can increase the perforin protein expression in the peripheral blood mononuclear cells in patients with CHB. The variation of perforin protein expression in the peripheral blood mononuclear cells may be closely related to the efficacy of interferon-alpha treatment against hepatitis B virus.

Adult ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Perforin ; metabolism ; Recombinant Proteins ; Treatment Outcome ; Young Adult

The purpose of this study was to observe whether human peripheral dervied monouncleas cells (hMNCs) could participate in the regeneration process of the ischemic hearts in the way of differentiating into cardiomyocytes, vascular endothelial cells and smooth muscle cells. hMNCs were transplanted into the bodies of the mice with myocardial infarction through the tail vein injection. Hearts were harvested 2-12 weeks after injection then sliced up into frozen sections of 5 micron thickness. Double immunofluorescence staining was used to test the differentiation of the grafted cells into cardiomyocytes, smooth muscle cells and vascular endothelial cells which revealed that cells expressing both HLA and TNT, HLA and alpha-SMA, HLA and vWF existed in the hearts of the mice. According to the study, it is probable that hMNCs could participate in the regeneration process of the infarcted hearts in the way of differentiation.
Animals ; Cell Differentiation ; physiology ; Humans ; Leukocytes, Mononuclear ; transplantation ; Mice ; Mice, Nude ; Myocardial Infarction ; pathology ; therapy ; Myocytes, Cardiac ; cytology ; Transplantation, Heterologous

Objective To discuss the clinical efficacy of internal fixation with miniature steel plate and kirschner pins in treatment for metacarpal neck fracture.Methods A total of 142 patients with cervix metacarpal fracture in our hospital from January 2015 to December 2016 were divided into the control group(n =71) and the observation group(n =71) randomly.The patients in control group were treated with internal fixation with kirschner pins,and the patients in observation group were treated by internal fixation with miniature steel plate.The operative time,postoperative healing time,postoperative pain,metacarpophalangeal joint function and complications were compared between the two groups.Results The operation time of observation group was (38.4 ± 3.8) minutes,the fracture healing time was (3.1 ± 1.4)weeks,the excellent and good rate of TAM was 97.2％ after operation,the excellent and good rate of FAFS was 98.6％ after operation,all these indexes were significantly better than those in the control group with statistical significance (P ＜ 0.05).Conclusion Internal fixation with miniature steel plate in the treatment of cervix metacarpal fracture has the advantages of exact clinical effect,short operative time,quick fracture healing and good recovery of joint function.

OBJECTIVETo investigate the role of CD4+CD25+high regulatory T cells in the pathogenesis of autoimmune hepatitis.

METHODSCD4+CD25+ high regulatory T cells and CD4+ T cells were measured by using flow cytometry in 16 patients with autoimmune hepatitis, 22 patients with chronic hepatitis B and 20 healthy blood donors. Foxp3 protein was detected by immunohistochemical assay in liver tissues from the patients with autoimmune hepatitis or chronic hepatitis B.

RESULTSThe percentage of CD4+CD25+high/CD4+ in patients with autoimmune hepatitis was significantly lower than that in healthy controls and patients with chronic hepatitis B. Meanwhile, the percentage of CD4+CD25+high/CD4+ highly increased in patients with chronic hepatitis B, compared with healthy controls; Foxp3 positive cells were mostly located in the hepatic lobular perisinusoidal spaces and the portal tract, and there was a significant difference in the quantity of Foxp3 positive cells between patients with autoimmune hepatitis and chronic hepatitis B.

CONCLUSIONPatients with autoimmune hepatitis harbor a decreased percentage of CD4+CD25+ high regulatory T cells, which may be associated with development of autoimmunity.

Adult ; Female ; Forkhead Transcription Factors ; analysis ; Hepatitis B, Chronic ; immunology ; Hepatitis, Autoimmune ; immunology ; Humans ; Immunohistochemistry ; Liver ; chemistry ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology

Objectives:To observe the clinical features and factorsrelated to treatment decision for hospitalizedpatients with mitral regurgitation (MR). Methods:A total of 3 450 consecutivepatients with transthoracic echocardiography (TTE) confirmed moderate to severe MR admitted in our hospital from 2014-01-01 to 2015-12-31 were enrolled. Base on therapeutic method, the patients were divided into 2 groups:Surgery group, n=1 845 and Medication group, n=1 605. The baseline data including TTE results were collected, clinical features were compared between 2 groups and factors related to treatment decision were analyzed. Results:Mean age of this patient cohort was (54.8±13.8) years including 26.99% (931/3 450) patients aged ≥65 years. The most common etiology was primary MR, 324 (9.39%) patients were asymptomatic at admission and decreased left ventricular ejection fraction (LVEF) was evidenced in 55.28% (1 907) patients. Total in-hospital mortality was 0.75% (26). Compared with Medication group, the patients in Surgery group were younger ([52.65±12.01] years vs [57.39±15.25] years), prevalence of severe MR (56.69% vs 26.79%) and primary MR (89.49% vs 39.00%), as well as LVEF value ([61.62±9.20] % vs [48.00±17.53] %) were higher (all P＜0.001).Logistic regression analysis indicated that age (OR=0.561, 95% CI 0.503-0.627), MR etiology (OR=3.062, 95% CI 2.565-3.654), MR grade (OR=0.103, 95% CI 0.085-0.126) and LVEF (OR=2.478, 95% CI 2.147-2.860) were the determinants for treatmentdecision making in hospitalized patients with moderate to severe MR. Conclusions:In this patient cohort, there are considerable proportion of aged patients with moderate to severe MR. Primary MRis the major etiology. 46.52% patients received conservative therpay instead of surgery, older age,secondary MR, moderate MR and decreased LVEF are the major reasons for choosing conservative therapy in this patient cohort.