Objective To explore whether arsenic trioxide(As2O3)-induced apopotosis in drug-resistant leukemia K562/ADM cells may induce in through endoplasmic reticulum stress leukemia cell apopotosis.Methods The apoptosis of K562/ADM cells was identified by double staining of FITC-Annexin V and propidium iodide(PI),the ultrastructure of the cells,endoplasmic reticulum and mitochondria were observed by transmission electron microscopy.Flow cytometry(FCM) was employed to assess mitochondrial inner membrane potential(??m),intracellular calcium concentration,cytochrome c(Cyt c) release and caspase-3 activity.The expression of GRP78 protein was analyzed by Western blot.Results During the apoptotic process of K562/ADM cells induced with 2 ?mol/L and 5 ?mol/L As2O3,the endoplasmic reticulum exhibited obvious expansion and degranulation,and the mitochondria illustrated inner and outer membranes fusion,reduced and confused cristae,swelling and vacuolization.The mitochondrial ??m decreased,the intracellular calcium concentration and releasing of cytochrome c from mitochondria increased,and caspase-3 was activated.Western blot result indicated upregulation of GRP78 protein at endoplas-mic reticulum in apopototic K562/ADM cells.Conclusion As2O3 can initiate the endoplasmic reticulum stress in K562/ADM cells,and induces to apoptosis of the drug-resistant cell via endoplasmic reticulum-mitochondrial pathway.

Objective To investigate the role and mechanism of RSPO1 in osteoblasts differentiation.Methods The xCELLigence system was used to study the toxicity and role of RSPO1 on the C2C12 cells proliferation.Alkaline phosphatase (ALP) activity was measured by using a phosphatase detection kit.The expression of osteoprotegerin (OPG) was determined using enzyme-linked immunosorbent assay (ELISA).Wnt/β-catenin signaling was evaluated using the TOPflash T cell factor (TCF) luciferase reporter assay.Western blotting assay was performed to confirm the accumulation of β-catenin protein.T test was used for statistical analysis.Results RSPO1 had no effect on the C2C12 cells proliferation,and it produced no toxicity to C2C12 cells.RSPO1 alone resulted in a slight increase in the activity of ALP (2.85±0.08 vs 1.74±0.21,t=3.014,P＜0.05) and the expression of OPG (1.29±0.13 vs 1.00±0.21,t=3.348,P＜0.05),whereas the combination of RSPO1 and Wnt3A significantly amplified ALP activity (81.37±5.08 vs 1.74±0.21,t=31.31,P＜0.01) and OPG protein expression (5.26±0.60 vs 1.00±0.21,t=6.99,P＜0.01).RSPO1 synergized with Wnt3A to activate TCF activity and induce accumulation of β-catenin (3.28±0.18 vs 1.00±0.21,t=10.94,P＜0.05).However,RSPO1 alone had no effect on the TCF activity and β-catenin accumulation (1.25±0.08 vs 1.00±0.21,t=2.225,P＞0.05).Conclusion Our study has revealed that RSPO1 synergized with Wnt3A in activating Wnt/β-catenin signaling pathway to induce osteoblasts differentiation,which demonstrate the therapeutic potential of RSPO1 for RA.

Objective To investigate the expression of anionic site in adriamycin-induced nephropathy (AN) rats, and to further explore the effects of Qufengtongluo Recipe on charge barrier in glomerulus in AN rats. Methods Adriamycin nephropathy was induced by a single tail intravenous injection of adriamycin. Totally 80 rats were randomly divided into normal control group and nephropathy model group. Three weeks later, the nephropathy model was established, and 50 AN rats were randomly divided into five groups: nephropathy model group (B, n=10), Qufeng group (C, n=10), prednisone and Qufeng group (D, n=10), prednisone group (E, n=10) and benazepri group (F, n=10), and they were treated respectively. With treatment being given respectively, renal tissue samples in each group were collected at week 3 and 7, respectively. The ultrastructure and expression of anionic site were examined by electron microscope observation. Results ① After adriamycin injection, a significant increase of the 24-hour urinary protein was observed at week 3 (P<0.01). In AN rats, serum albumin was decreased (P<0.01) while serum TCH and TG were increased (P<0.01). ② In AN rats the diffuse fusion and effacement of foot processes and decrease of anionic sites in GBM were observed at week 3. ③ At week 7, the average intensity of AS dramatically increased in C and E groups (P<0.01) compared with that in nephropathy model group. Conclusion The abnormal expression of AS is the important mechanism that leads to the occurrence and development of proteinuria in AN rats. It is possible that Qufengtongluo Recipe has effects on nephrotic syndrome through altering the charge barrier in GBM in glomerulus.

Objective To investigate the expression of nephrin mRNA in adriamycin-induced nephropathy (AN) in rats, and to explore the effect of Qufengtongluo recipe on proteinuria in AN rats and on the expression of nephrin mRNA. Methods Adriamycin nephropathy was induced by a single tail intravenous injection of adriamycin. We randomly divided 140 rats into a normal control group (n=32) and a nephropathy model group (n=108). Three weeks later, 90 AN rats were randomly divided into 5 groups: a model group, a qufeng group, a qufeng and prednisone group, a prednisone group, and a benazepri group (18 rats in each group). They were treated respectively, and renal tissue samples were collected at week 0, 3, 5, and 7, respectively. The distribution and expression of nephrin mRNA were examined by indirect immunofluorescence and semi-quantity RT-PCR. Results In the AN rats, the diffuse fusion and effacement of foot processes were observed at week 3. The fluorescence intensity of nephrin and the expression of nephrin mRNA significantly increased in the qufeng group and the prednisone group compared with the model group at the week 7 (P＜0.01).Conclusion Abnormal expression of nephrin is the important molecular mechanism that leads to the occurrence and development of proteinuria in AN rats. Qufengtongluo recipe has effect on nephrotic syndrome through altering the expression and distribution of nephrin in glomerulus.

BACKGROUND:Studies have funded that reduced Wnt/β-catenin signaling is involved in the onset and/or progression of bone erosion in rheumatoid arthritis. It can lead to potential new treatment approaches of bone erosion by enhancing Wnt/β-catenin signaling pathway. R-spondin 1 may act as a Wnt agonist, but there is no study in human osteoblasts. OBJECTIVE:To verify the effect of R-spondin 1 on promoting differentiation and maturation of human osteoblasts by inhibiting DKK1. METHODS:S40-transfected human osteoblast lines, hFOB1.19, were treated with R-spondin 1, Wnt-3a and DKK1 to detecting the proliferation, alkaline phoshpatase activity and osteoprotegerin concentration. RESULTS AND CONCLUSION:R-spondin 1 had no effects on hFOB1.19 cel s. Wnt-3a upregulated the activity of alkaline phoshpatase, which could be enhanced by addition of R-spondin 1. R-spondin 1 could reduce the DKK1-mediated inhibition of alkaline phoshpatase activity in hFOB1.19 cel s. R-spondin 1 increased the concentration of osteoprotegerin, and moreover, the promotion of osteoprotegerin by R-spondin 1 alone was stronger than the inhibition by DKK1. These findings suggest that R-spondin 1 can inhibit DKK1 by Wnt/β-catenin signal pathway to promote the differential and maturation of human osteoblasts to excrete osteoprotegerin.

Objective To evaluate the efficacy of different duration of low-carbohydrate diet (LCD) intervention on glycosylated hemoglobin (HbA1c) in type 2 diabetes mellitus (T2DM) patients. Methods Randomized controlled trials (RCTs) of LCD intervention in T2DM patients were collected in the databases such as MEDLINE, PubMed, OVID, China National Knowledge Infrastructure, China Scientific Journal Database by VIP, Wanfang database, et al.Data were analyzed by RevMan 5.3 version. Results Eight RCTs were included. The results of Meta-analysis indicated that the effects of lowering HbA1c by LCD intervention for three (Z=2.28, P<0.05) and six months (Z=14.99, P<0.01) were better than other diabetes diets, but there was no significant statistical difference between one (Z=0.65, P=0.51) and two years (Z=1.62, P=0.10). Conclusions Hypoglycemic effect of short-term LCD was better than other diabetes diets, but long-term effect was similar between them. LCD was a therapeutic diet suitable for T2DM patients.

Objective To explore the clinical feature and gene types in patients with primary carnitine deficiency. MethodsClinical data of 6 patients with primary carnitine deficiency and 2 patients with maternal carnitine deficiency found in the screening by tandem mass spectrometry technology during December 2013 to December 2016 were retrospectively analyzed. Results The free carnitine levels of 8 patients in initial and recall screening were 5.85±1.65 μmol/L and 5.22±1.02 μmol/L. Two pathogenic alleles were detected in each patient with primary carnitine deficiency by genetic and metabolic disease panel based on Ion Torrent semiconductor sequencing. After treatment with oral L-carnitine, the free carnitine levels of 6 patients with primary carnitine deficiency were 20.24±3.88 μmol/L. The carnitine levels returned to normal after mixed feeding for one week in 2 patients with maternal carnitine deficiency, and no genetic diagnosis was carried out. Conclusion Primary carnitine deficiency can be effectively detected using tandem mass spectrometry technology and next generation sequencing panel and the prognosis is good with early standard treatment.

Background:Chronic constipation is a major cause of impaired quality of life in modern society. Reasonable and effective management of chronic constipation could be achieved based on the principle of evidence-based medicine and the modern concept of constipation,and this is a challenge facing the clinicians. Aims:To investigate the role of barium-based colonic transit detection in diagnosis and treatment of chronic constipation. Methods:Fifty patients with chronic constipation from Apr. 2013 to Oct. 2014 at the First Hospital of Shanxi Medical University were recruited and randomly allocated into two groups,control group and individualized treatment group. Patients in individualized treatment group received 20 barium markers orally and abdominal plain radiography was performed 48 and 72 hours later,respectively for calculating the colonic transit index. According to the type of colonic transition and the characteristics of colonic motility estimated by colonic transit index and clinical manifestations,an individualized therapeutic regimen was formulated and the efficacy was evaluated. Patients in control group were treated empirically according to the clinical manifestations. Results:Mosapride and lactulose or polyethylene glycol were administered orally in control group;when abdominal pain or abdominal distension was predominant,pinaverium bromide or trimebutine was used respectively instead of mosapride. Barium-based colonic transit detection revealed that 9 patients in individualized treatment group were slow transit constipation,6 were outlet obstructive constipation and 8 were the mixed type. After 2 weeks of empirical or individualized treatment,the defecation rates of the two groups were 24. 0%(6 / 25)and 52. 2%(12 / 23)within 24 hours and 64. 0%(16 / 25)and 87. 0%(20 / 23)within 48 hours,respectively(P all < 0. 05). Conclusions:Barium-based colonic transit detection is a simple,economical and practical modality for guiding the individualized treatment in patients with chronic constipation.

Objective To explore the significantly differentially.expressed microRNAs during antibody-mediated renal allograft rejection.Method MicroRNA array assay was used,and the obtained data were analyzed by bioinformatics analysis.The obtained significant microRNAs were further analyzed to forecast the targeted genes in the common database,then experimental means were used to testify the targeted genes.Result During the antibody-mediated renal allograft rejection,the significantly over-expressed microRNAs were miR-200c,miR-200b,miR-30c,miR-30b and miR-30e+,etc.The significantly down-expressed microRNA was miR-338-5p.The bioinformatics analysis results indicated that TRAF3 was the targeted gene of miR-338-5p,which was testified by real time PCR,immunohistochemical assay and fluorescence reporter assay.Conclusion miR-338-5p anticipated in the antibody-mediated renal allograft rejection by targeting TRAF3.

To explore the serum miR-338-5p expression characteristics in renal transplant recipients ,and the role of regulating BAFF signal ,then investigate its biological significance.Methods:Healthy volunteers were enrolled as control group.Serum miR-338-5p was detected by real-time PCR;soluble BAFF was detected by ELISA;anti-HLA-Ⅰantibody,anti-HLA-Ⅱ antibody and anti-MICA antibody were detected by liquid chip technology.SPSS17.0 software was applied.t-test was used to compare the means of two independent samples;Paired samples t-test was used to compare the means of two paired samples;Spearman method and Pearson method were used to analyse the correlation;P<0.05 was considered to be statistically significant.Results: Compared with healthy controls,serum miR-338-5p in renal transplant recipients decreased significantly (P<0.001),while serum BAFF increased significantly (P<0.01).Serum miR-338-5p levels within 1 year post-transplantation were significantly lower than that of more than 1 year post-transplantation (P<0.01);Serum miR-338-5p levels within 3 years post-transplantation were significantly lower than that of more than 3 years post-transplantation (P<0.01);To all research objects,serum miR-338-5p was significantly negatively correlated with serum BAFF (r=-0.51,P<0.001),and serum miR-338-5p was significantly negatively correlated with anti-HLA-Ⅱ antibody(r=-0.322, P<0.05);Serum miR-338-5p within 3 years was significantly negatively correlated with anti-HLA-Ⅱantibody (r=-0.423,P<0.05), and serum miR-338-5p within 3 years was significantly negatively correlated with anti-MICA antibody(r=-0.411,P<0.05);Serum miR-338-5p more than 3 years was significantly positively correlated with anti-MICA antibody(r=0.486,P<0.05),and Serum miR-338-5p more than 3 years was significantly positively correlated with anti-HLA & MICA antibody(r=0.578,P<0.01).Conclusion:miR-338-5p may directly or indirectly target BAFF signal ,and participate antibody mediated immune response by regulating its target genes and interfere with the long-term survival of transplanted renal.