Osteoclastic bone resorption and osteoblastic bone formation are coordinated as a coupled mechanism to effect the development of bone and to maintain bone homeostasis. Recently reported Eph/ephrin bidirectional signaling between osteoclasts and osteoblasts plays a pivol role in bone homeostasis and casts new light on coupling of bone resorption and bone formation, which is gaining more and more attention in researches of bone biology and bone diseases. The present article aims to address the researches on the Eph/ephrin bidirectional signaling between osteoblasts and osteoclasts with molecular constitution, mechanism of the signal transduction, biological significance and so on.

OBJECTIVETo construct Dishevelled 2 (Dvl2)-targeted siRNA plasmids and to identify the effective recombinant plasmids in transciently-transfected RAW264.7 cells.

METHODSThe interfering sequences of Dvl2 were designed according to the sequence of Dvl2 of GenBank. Five paires of oligonucleotides were synthesized and inserted into plasmid pMAGic 4.0 to generate siRNA expression vectors, which were identified by flora PCR and sequence analysis. The recombinant plasmids siRNA-Dvl2 was transciently transfected into RAW264.7 cells by Lipofectamine 2000, which was confirmed under a fluorescence microscope and the interfering efficiency was detected by real-time RT-PCR.

RESULTSFive Dvl2 siRNA frames were successfully inserted into the plasmid vector pMAGic 4.0, and the flora PCR and sequence analysis confirmed the correct construction. Three of the five siRNA vectors suppressed the expression of Dvl2 mRNA, in which the siRNA-Dvl2-3 was the most efficient.

CONCLUSIONThe Dvl2-targeted recombinant siRNA plasmids can be constructed successfully to inhibit Dvl2 mRNA expression in transciently-transfected RAW264.7 cells, which can be used to pack virus particles and to construct siRNA-Dvl2 stably-transfected RAW264.7 cells in further research.

Genetic Vectors ; Humans ; Plasmids ; RNA, Messenger ; RNA, Small Interfering ; Transfection

AIM:To investigate the effects of grasp seed procyanidins(GSP) on homocysteine-induced proliferation and migration in vascular smooth muscle cells(VSMC) and related molecular mechanisms.METHODS: Cell count and -TdR assay were used for detecting cell proliferation and DNA synthesis,ELISA assay was used for detecting inflammatory response,DCFH-DA assay for examining the levels of reactive oxygen species(ROS),Western blotting for detecting protein expression.RESULTS: Homocysteine(0.1-1 mmol/L) increased VSMC proliferation and migration,and the levels of ROS were in a dose-dependent manner.The results of Western blotting showed that homocysteine significantly increased the expression of MCP-1,IL-6 and TNF-?.However,Compared with control group,in GSP(5-20 g/L) group,the increased VSMC proliferation,migration and the production of ROS and the expression of MCP-1,IL-6 and TNF-? mediated by homocysteine were markedly suppressed.EMSA showed that in GSP treatment group,the NF-?B activation was also almost completely inhibited.CONCLUSION: GSP inhibits homocysteine-induced VSMC proliferation,migration and inflammatory response through interfering with ROS dependent on NF-?B signal pathway.

Objective To describe the clinical characteristics of idiopathic ventricular fibrillation (IVF) with fragmented QRS complex (f-QRS) and J wave in resting electrocardiogram. Methods We reviewed data from 21 case subjects in our hospital who were resuscitated after cardiac arrest due to IVF and assessed the prevalence of f-QRS and J wave in resting electrocardiogram (ECG). All the case subjects were classified among three groups based on the electrocardiographic morphology: group I, both f-QRS and J wave were observed (n = 6), group II, only J wave was observed (n = 9), group III, neither f-QRS nor J wave was observed (n = 6). Population characteristics, history of syncope or sudden cardiac arrest, incidence of ventricular fibrillation (VF), and circumstance of VF were evaluated among the three groups. Results The incidence of index events (syncope, survived cardiac arrest and VF episodes recorded in implantable cardioverter defibrillator (ICD) or pacemakers) was 13.4 ± 5.6 per-year in group I, 10.8 ± 3.9 per-year in group II, and 9.8 ± 4.2 per-year in group III. There were significant differences in incidences among the three groups, the most frequent index events were observed in group I. The hazard ratio for incidence was 3.2 (95%CI, 1.1-7.9; P = 0.01). The history and circumstance of the index events were different among the groups. In group I, all the index events occurred during sleep in early morning. In group II, four subjects suffered VF during strenuous physical activities or agitation state, two during sleep in early morning, three in usual activity. In group III, one subject suffered VF during sleep in early morning, one in agitation state, four in usual activity. Conclusions This study suggests that the IVF patients with the combined appearance of f-QRS and J wave in the resting ECG suffer an increased risk of VF, this subgroup of IVF patients has a unique clinical feature.

OBJECTIVETo explore the clinical effect of L-ornithine L-aspartate (LOLA) granules in treating chronic liver disease in patients with high-level serum gamma-glutamyltransferase (G-GT) using a 24-week treatment course.

METHODSTwo-hundred patients with chronic liver disease and above normal G-GT were given a 12-week course of LOLA granules (9 g/d) and then classified into the following three groups according to the change in serum Gamma-GT:group I:patients with Gamma-GT level returned to normal;group II:patients with serum Gamma-GT level that was reduced during the treatment; group III:patients with serum Gamma-GT level that did not decrease or that increased to a higher level than at start of treatment.After the 12-week treatment course, the patients in group I were divided into three subgroups for receipt of a control drug (compound glycyrrhizin, 50mg/d) or an additional 12-week course of Gamma-GT at a reduced dose (LOLA granules 3 g/d) or at the original dose; groups II and III were maintained on the initial dose for an additional 12 weeks.The groups were reassessed at the end of the second 12-week course (at the end of week 24 of the study's observation period).Count data were compared using the x2 test and measurement data were compared using the t-test.

RESULTSIn group I, the serum Gamma-GT level was 90.9% at the end of the first 12-week course and dropped to a mean level of 52.2% for both of the subgroups that received the reduced and original dose after the additional 12 weeks of LOLA granules treatment; the difference from week 12 to week 24 was significant (x2=8.213, P less than 0.05).The 24-week change in serum Gamma-GT levels for the group I reduced and original dose subgroups vs.the control subgroup were also significantly different from those seen in groups II and III (P less than 0.05).The percentage of patients in group I who achieved normal level serum Gamma-GT after 24 weeks of treatment (78.6%) was significantly higher than that for the control group (vs.55.0%, x2=11.452, P less than 0.05).When the patients in group 1 who had received the 12 additional weeks of LOLA granules treatment were measured again at two weeks after the treatments had been discontinued (end of week 26), the percentage of patients with normal serum Gamma-GT level was 92.7%, with only three cases showing obviously abnormal levels; in contrast, the group I patients in the control group of the second 12-week study period had on 66.7% of patients with normal-level serum Gamma-GT.The difference in change between the treated groups (both reduced and original dose) and the control group was significant (x2=14.964, P less than 0.05).

CONCLUSIONPatients whose serumGamma-GT levels returned to normal after receipt of LOLA granules for 12 weeks benefitted from an additional 12 weeks of consolidation treatment, and those given the treatment at the original dose benefitted most.Compared with the compound glycyrrhizin, LOLA granules provided a better maintenance of resolved Gamma-GT level.Therefore, the effect of LOLA appears to be reliable and stable as well as safe for clinical use.

Chronic Disease ; Dipeptides ; therapeutic use ; Humans ; Liver Diseases ; drug therapy ; Liver Function Tests ; gamma-Glutamyltransferase ; blood

Objective To understand the incidence and death patterns of viral hepatitis in China and provide evidence for the prevention and control of viral hepatitis.Methods The analysis was conducted on the incidence and death data of viral hepatitis published by National Health and Family Planning Commission during 2004-2013.Results The incidences of viral hepatitis in Guizhou,Yunnan,Tibet,Gansu,Qinghai,Ningxia and Xinjiang provinces (autonomous region) were high.The major forms were hepatitis B (80.63/100 000) and hepatitis C (9.68/100 000),accounting for 80.90％ and 9.25％ of the total reported viral hepatitis cases respectively.The incidences of hepatitis A and unidentified hepatitis decreased and the incidence of hepatitis B,C and E increased during this period.During the 10 years,10 008 deaths caused by viral hepatitis were reported (1 001 deaths per year).The reported deaths caused by hepatitis A,hepatitis E and unidentified hepatitis decreased during this period.The reported deaths caused by hepatitis B were in a downward trend,but the constituent in total cases remained high.The reported deaths caused by hepatitis C were in an upward trend.Conclusion During 2004-2013,the overall incidence of viral hepatitis showed no downward trend in China.The incidence of hepatitis B remained high,and the incidence of hepatitis C showed an obvious upward trend.The overall death rate and case fatality rate of viral hepatitis showed a downward trend,but hepatitis B remained the main cause of viral hepatitis related death,and the death caused by hepatitis C was in increase.Hepatitis B and hepatitis C are the major targets in the prevention and treatment of viral hepatitis in China,and the 7 western provinces (autonomous region) with high incidences are the key regions of the prevention and control.

ObjectiveTo explore the possible mechanisms of Tongfengning （痛风宁， TFN） in treating hyperuricemia （HUA） of spleen deficiency with exuberance of dampness syndrome. MethodsTen of 60 mice were randomly selected， and were fed with regular diet as the control group， while the remaining 50 mice were fed with high-fat and high-sugar diet combined with excessive exercise and potassium oxonate-allopurinol suspension to establish an HUA animal model of syndrome of spleen deficiency with exuberance of dampness. After the successful modeling， in order to better observe the effects of TFN on the intestinal microbiota of the model mice， a mixed antibiotic suspension was administered by gavage to induce further dysbiosis of the intestinal microbiota in the model mice. Fifty sucessfully modeled mice were randomly divided into model group， TFN group， allopurinol group， probiotics group， and an allopurinol + probiotics group， 10 in each group. The TFN group was administered TFN liquid at a dosage of 19.11 g/（kg·d） by gavage. The allopurinol group was administered allopurinol suspension at a dosage of 78 mg/（kg·d） by gavage. The probiotics group was administered live combined Bifidobacterium and Lactobacillus tablets suspension at a dosage of 3 g/（kg·d） by gavage. The allopurinol + probiotics group was administered allopurinol at a dosage of 78 mg/（kg·d） and live combined Bifidobacterium and Lactobacillus tablets suspension at a dosage of 3 g/（kg·d） by gavage. The control group and model group were administered normal saline at a dosage of 19.11 ml/（kg·d） by gavage. The interventions were continued for 21 days. In order to maintain a stable high blood uric acid state， all groups but the control group continued modeling while receiving drug intervention. The changes in spleen deficiency syndrome scores， blood uric acid levels， microbial community structure， acetic acid and butyric acid content in intestinal lavage fluid， adenosine deaminase （ADA） and xanthine oxidase （XOD） content in small intestine tissue， as well as ATP-binding cassette transporter G2 （ABCG2）， glucose transporter 9 （GLUT9） protein and mRNA expression in the small intestine tissue were compared among the groups of mice. ResultsCompared with the control group， the model group showed increased spleen deficiency syndrome scores， blood uric acid levels， relative abundance of phylum Firmicutes， Firmicutes/Bacteroidetes ratio， abundance of Bacteroides genus， Klebsiella genus， and Enterococcus genus， acetic acid content in intestinal lavage fluid， ADA and XOD content in small intestine tissue， as well as GLUT9 protein and mRNA expression （P<0.05）. The number of operational taxonomic units （OTUs） of intestinal microbiota， relative abundance of Bacteroidetes phylum， abundance of Lactobacillus genus and uncultured Bacteroides genus， butyric acid content in intestinal lavage fluid， and ABCG2 protein and mRNA expression in small intestine tissue were significantly decreased （P＜0.05）. Compared with the model group， in the group treated with TFN， probiotics， and allopurinol + probiotics， the spleen deficiency syndrome score， blood uric acid level， relative abundance of Firmicutes， acetic acid content in intestinal lavage fluid， ADA and XOD content in small intestine tissue， GLUT9 protein and mRNA expression significantly decreased. The number of gut microbiota OTUs， relative abundance of proteobacteria， butyric acid content in intestinal lavage fluid， ABCG2 protein and mRNA expression in small intestine tissue significantly increased （P＜0.05）. In the probiotics group， the ratio of Firmicutes to Bacteroidetes decreased. In the TFN group， the abundance of Lactobacillus and uncultured Bacteroidetes significantly increased， while the abundance of Parabacteroides， Klebsiella， and Enterococcus significantly decreased （P＜0.05）. Compared with the TFN group， allopurinol group and the probiotics group showed elevated blood uric acid levels， abundance of Bacteroidetes， ADA and XOD levels in intestinal tissue， and GLUT9 mRNA expression. The relative abundance of Firmicutes， abundance of lactobacilli， and ABCG2 mRNA expression significantly decreased. The probiotics group showed elevated GLUT9 protein expression in intestinal tissue. The probiotics group and the allopurinol plus probiotics group showed significantly higher scores for spleen deficiency syndrome in mice， and lower levels of butyric acid in mouse intestinal lavage fluid. The allopurinol group showed decreased numbers of OTUs in mouse intestinal flora， decreased abundance of proteobacteria， and butyric acid levels in intestinal lavage fluid. The allopurinol group also showed decreased ABCG2 protein expression in intestinal tissue， increased acetic acid levels in intestinal lavage fluid， increased abundance of Klebsiella， and significantly elevated GLUT9 protein expression （P＜0.05）. ConclusionsThe treatment of HUA with TFN may be associated with the regulation of intestinal probiotics （such as lactobacilli） and pathogenic bacteria （such as Klebsiella）， as well as the production of bacterial metabolites such as acetic acid and butyric acid. It may also involve reducing the expression of ADA and XOD in the intestines， decreasing intestinal uric acid production， upregulating the expression of intestinal epithelial urate transporter ABCG2， downregulating GLUT9 expression， and promoting intestinal uric acid excretion. These factors are related to the syndrome of spleen deficiency with exuberance of dampness.

To explore the pharmacogenomic markers that affect the platinum-based chemotherapy response in non-small-cell lung carcinoma (NSCLC), we performed a two-cohort of genome-wide association studies (GWAS), including 34 for WES-based and 433 for microarray-based analyses, as well as two independent validation cohorts. After integrating the results of two studies, the genetic variations related to the platinum-based chemotherapy response were further determined by fine-mapping in 838 samples, and their potential functional impact were investigated by eQTL analysis and in vitro cell experiments. We found that a total of 68 variations were significant at P < 1 × 10^-3 in cohort 1 discovery stage, of which 3 SNPs were verified in 262 independent samples. A total of 541 SNPs were significant at P < 1 × 10^-4 in cohort 2 discovery stage, of which 8 SNPs were verified in 347 independent samples. Comparing the validated SNPs in two GWAS, ADCY1 gene was verified in both independent studies. The results of fine-mapping showed that the G allele carriers of ADCY1 rs2280496 and C allele carriers of rs189178649 were more likely to be resistant to platinum-based chemotherapy. In conclusion, our study found that rs2280496 and rs189178649 in ADCY1 gene were associated the sensitivity of platinum-based chemotherapy in NSCLC patients.