BACKGROUND:Bone turnover markers are the products in the blood or urine in the process of bone reconstruction, which can be used to evaluate bone resorption and bone formation rate, thereby indicating potential bone diseases. OBJECTIVE:To analyze the effect of exercise on the levels of bone turnover markers in the blood and urine samples. METHODS:Databases of PubMed and Wangfang were retrieved with key words of“bone formation, bone resorption, alkaline phosphotase, osteocalcin PICP, PINP, hydroxyproline, pyridinoline, tartrate resistant acid phosphatase”by screening titles and abstracts to search papers related to exercise effects on the biological markers of bone formation. Final y, 48 papers were discussed. RESULTS AND CONCLUSION:The effect of exercise on bone turnover refers to the influence on bone formation and bone resorption. Exercise enhances or inhibits the activity of osteoblasts and osteoclasts, and accelerates or delays bone reconstruction by promoting or inhibiting the growth of osteoblasts and osteoclasts. In recent years, exercise exhibits more and more influence on the bone. Bone turnover markers compared with bone mineral density show changes earlier. By measuring the levels of bone turnover markers in blood and urine samples, it help us to understand the metabolism of bone tissue, to evaluate bone metabolic state, osteoporosis diagnosis classification, predicting of fracture risk, to observe the curative effect of drug treatment, and to diagnosis the metabolic bone differential disease.

The process of exercise regulating bone metabolism is complicated, which involves a number of signaling pathways. A large number of studies in vitro have indicated that mechanical stress regulates bone metabolism by Wnt, bone morphogenetic protein (BMP), and osteoprotegerin (OPG)/receptor activator of NF-κB ligand (RANKL)/receptor activator of NF-κB (RANK) signaling pathways. Both the in-tensity and frequency of mechanical stress have varing impact on bone tissue and cells. Plenty of studies in vivo also have shown that exer-cise regulates bone metabolism by key factors in bone metabolism signaling pathways. This paper reviewed the effects of exercise on bone metabolism pathways and their mechanisms.

Objective:3D hydrogel cell model was established,and cyclic compressive loading on MC3T3-E1 cell with different intensities,frequencies and durations was applied,in order to research the suitable solution about promoting the osteoblast differentiation with cyclic compression.Methods:Cyclic compressive loading on MC3T3-E1 cell was applied with different intensity,frequency and time.After compressive loading finished,the total RNA extraction from cell-gel constructs were performed and quantified ATF4,ALP,Runx2,Osteocalcin,RANKL and RANK mRNA.Results:RANKL and RANK mRNA expression significantly with different frequencies cyclic compressive loading (P ＜ 0.05),and ALP mRNA (P ＜ 0.05) and Runx2 mRNA (P ＜ 0.01) expression significantly with different intensities and frequencies cyclic compressive loading (P ＜ 0.05).Meanwhile,Runx2 mRNA expression with 4h significant higher than 12h (P ＜ 0.05),and RANKL mRNA expression with 4h significant lower than 12h (P ＜ 0.05).Conclusion:Determine the stress intensity and frequency,1％ intensity,frequency of 0.5 Hz,4 h of cyclic compression intervention could promote the growth of osteoblasts-like cells in the 3D hydrogel model.

Objective To study early diagnosis and treatment of Wernicke's encephalopathy(WE) in allogeneic peripheral blood stem cell transplantation recipients.Method A 17 years old patient with acute B-lymphocytic leukemia received HLA-matched nonrelative allogeneic peripheral blood stem cell transplantation after conditioning with total-body irradiation/idamycin/cyclophosphamide (TBI/IDA/Cy) regimen.CD25 monoclonal antibody and cyclosporine A+mycophenolate mofetil + methotrexate were administrated for graft versus host disease prophylaxis.Result On the day 8,the platelet was over 20 × 109/L; On the day 10,the neutrophile granulocyte was over 0.5 × 109/L; On the day 28,full engraftment was confirmed by a bone marrow medicolegal identification.The continued nausea and vomiting after HSCT resulted in deficiency of intake and malabsorption.On the day 54,illusion and tremor occurred,and the follow-up brain MRI suggested WE,but the patient died before thiamine replacing therapy.Conclusion WE is also a rare neurologic complication of HSCT,however,it can easily be overlooked.So early radiologic surveillance and treatment for patients with WE is very important to minimize central nervous system complications and unwanted mortality.

Objective To develop a liquid chromatography technique coupled with tandem mass spectrometry(LC-MS/MS)for simultaneous determination of four active catechins EGCC,ECG,EGC,and EC of tea polyphenols(TP)in rat plasma in order to further study its multi-component pharmacokinetics.Methods Following a single step liquid-liquid extraction of plasma samples with ethyl acetate,the four catechins were separated on a Hypersil ODS C18 column using an isocratic mobile phase composed of methanol-water(30:70).The detection using a mass spectrometer was performed under negative ESI in the MRM mode.The analytes were identified by reference to both MRM and tR values and quantified using peak area internal standard method.Results The method was shown to be specific without interference from matrix,metabolites,and impurities present in TP raw material and to be sensitive with LOD and LOQ of 1.5 and 10 ng/mL(EGCG)as well as 0.75 and 5 ng/mL(ECG,EGC,and EC).A good linearity was obtained over a wide range of 10-10000 ng/mL for EGCG and 5-5000 ng/mL for other three catechins(r > 0.996).The method was validated to be reproducible and reliable,as evidenced by intra-batch and inter-batch precision of less than 10% and 11%,accuracy of 97.13%-106.05% and 99.22%-103.14%,respectively.The recovery of extraction ranged from 72.74% to89.13%,matrix effect from 88.76% to 105.97% for four cateckins.The method was successfully used to study the pharmacokinetics of TP iv administered to rats at a dose of 100 mg/kg.Conclusion This method is shown to completely meet requirements for the multi-component pharmacokinetic study of TP in rats.

LC-MS/MS method was used to simultaneously determine anti-oxidative active catechins EGCG, ECG, EGC and EC in plasma of rats treated with tea polyphenols (TP). The integrated plasma concentration (C') of TP was calculated by means of self-defined weighing coefficient based on percent AUC of individual components, thereby assessing integrated pharmacokinetic (PK) parameters of TP via log C'-T curve. The anti-free radical effects of TP were estimated using inhibitory rate of drug-containing serum collected at different times from rats against in vitro lipid peroxidation of mouse liver homogenate. The obtained E-T curves were used to calculate anti-free radical pharmacodynamic (PD) parameters of TP. E-logC and E-log C' plots and linear regression were carried out in order to obtain the correlation coefficient (R2). The results indicated that the log C'-T curves of TP, which could be best described by three-compartment model, corresponded to elimination rule of iv administration of drugs. The integrated PK parameters showed that TP was distributed in body rapidly and widely, and eliminated from deep compartment slowly. From comparison of R2 values and consistence of C'-T course and E-T course, it was evident that TP integrated PK behaviors correlated much better with its PD behaviors than individual active components, and thus demonstrated that integrated PK parameters could characterize to maximal extent holistic disposition of Chinese herbal drugs and reflect residence properties of holistic effective substances in biological body.

This article is report the study of the pharmacokinetics and metabolic disposition of exogenous phosphocreatine (PCr) in rats by means of an ion-pair HPLC-UV assay. PCr and its metabolite creatine (Cr) and related-ATP in rat plasma and red blood cell (RBC) were simultaneously determined. A blank plasma and RBC were initially run for baseline subtraction. Plasma and RBC samples were deproteinized with 6% PCA prior to HPLC. Following i.v. administration of PCr 500 mg x kg(-1) and 1 000 mg x kg(-1) the C-T curve could be described by the two-compartment model with t1/2beta 22.5-23.3 min, V(d) 0.956 4-0.978 6 L x kg(-1), CL 0.029 L. kg(-1) x min(-1). The Cr as PCr degraded product appeared as early as 2 min post i.v. dosing with t(max) 20 min, t1/2kappa (m) 40.6-42.7 min and f(m) 60%-76%. After po administration of PCr, the parent drug in plasma was undetectable, but the metabolite Cr was detected with t(max) 65-95 min, t1/2kappa (m) 56.0-57.7 min, metabolite-based bioavailability F(m) 55.02%-62.31%. PCr i.v. administration resulted in significant elevation of ATP level in RBC but not in plasma, the related-ATP in RBC was characterized by t(max) 68-83 min, t1/2kappa 49-52 min. In RBC no exogenous PCr was found but Cr was detected following i.v. administration of PCr, with the t(max) 120 min and t1/2k (m) 70 min for Cr. The above results indicate that PCr eliminates and bio-transforms in body very rapidly; K > K(m) confers ERL, instead of FRL, type upon the metabolic disposition of Cr. Following po administration of PCr, the degraded product Cr is absorbed but not the parent drug PCr. The formed Cr can be accounted for by most of i.v. and po PCr. Intravenous dosing leads apparently increased and sustained Cr and related-ATP concentration in RBC.

Objective: To analyze the association between the concentration of metalloestrogens ( MEs ) in seminal plasma and sperm quality in infertile patients, so as to provide the basis for the study of MEs on human sperm quality. Methods: The spermatozoa concentration, progressive rate ( PR ), normal morphology rate ( NMR ) and DNA fragmentation index ( DFI ) were determined in the infertile male patients from Zhejiang Provincial People's Hospital from March to April, 2020. The contents of As, Cd, Co, Cr, Cu, Hg and Ni in seminal plasma were determined by inductively coupled plasma mass spectrometry ( ICP-MS ). The distribution of MEs in seminal plasma and the sperm quality in different concentration of MEs ( grouped by quartiles ) were analyzed. Results: Among 105 cases recruited, 28 cases were normal in sperm parameters and 77 cases were abnormal, including 22 cases of oligospermia, 47 cases of asthenospermia, 54 cases of dysspermia and 30 cases of abnormal DFI. As, Cr, Cu, Hg and Ni were detected in all samples, the detection rates of Cd and Co were 78.57%-90.91%. Compared with the normal group, the concentration of As in oligospermia group was higher [( 18.96±12.56 ) µg/L vs. (13.67±4.19) µg/L, P<0.05],the concentration of Cr was higher in asthenospermia, dysspermia and abnormal DFI in Q3 and Q4 groups groups [( 386.62±81.92 ), ( 378.02±81.46 ), ( 393.88±77.03 ) µg/L vs. ( 343.12±55.08 ) µg/L,P<0.05]. The PRs in Q3 and Q4 groups of Cr were lower than that in Q2 group [( 32.95±18.22 )%, ( 27.74±22.77 )% vs. ( 54.18±24.64 )%, P<0.05], the DFI in Q3 and Q4 groups was higher than that in Q2 group [( 26.91±14.77 )%, ( 29.91±16.93 )% vs. ( 9.87±10.93 )%, P<0.05], and the NMR in Q4 group was lower than that in Q2 group [( 1.62±1.72 )% vs. ( 3.36±1.97 )%, P<0.05]. The concentration of sperm in Q3 group of Cu was higher than that in Q2 group [( 115.87±88.22 )×106/mL vs. (61.91±66.16)×106/mL, P<0.05]. Conclusion As and Cu in seminal plasma are associated with abnormal sperm concentration, and Cr is associated with NMR, PR and DFI.

Objective: To create a model for early prediction of essential hypertension (EH) based on the TreeNet algorithm, so as to provide a tool for early monitoring of EH. Methods: The health examination data were collected from individuals receiving health examinations in Hangzhou Haiqin Health Examination Center or Shanghai Yibao Health Management Co., Ltd from 2014 to 2016, and a predictive model for EH was created based on the TreeNet algorithm. The effectiveness of the model for early prediction of EH was evaluated using root mean square error (RMSE), mean absolute deviation (MAD), coefficient of determination (R2) and receiver operating characteristic (ROC) curve. Results: A total of 12 variables were included in the model, and the highest contributing variable was body mass index (BMI), followed by BMI difference, two-year BMI difference, two-year triglyceride (TG) difference, two-year total cholesterol (TC) difference, high-density lipoprotein cholesterol (HDL-C) in 2014, TG in 2014, low-density lipoprotein cholesterol (LDL-C) in 2014, body weight in 2015, fasting blood glucose in 2015, TG in 2015, urea nitrogen difference and platelet in 2015. The highest predictive accuracy was 100.00%, and the lowest was 56.89%. The risk of EH significantly increased among individuals with BMI in 2015 of >25 kg/m2, two-year BMI difference of >0.5 kg/m2, two-year TG difference ranging from 1.3 to 3.3 mmol/L, TC in 2015 of 2.0 to 2.4 mmol/L and HDL-C in 2014 of <0.52 mmol/L. The model presented RMSE of 0.082, MAD of 0.064, R2 of 0.811, area under the ROC curve of 0.788 (95%CI: 0.741-0.815), sensitivity of 69.05% and specificity of 66.21% for prediction of EH Conclusion The TreeNet algorithm-based model is effective for early monitoring of high-risk individuals for EH.

OBJECTIVETo clarify the regulation of p53 through telomere pathway by investigating the molecular interaction between p53 and the main telomere-associated protein Telomeric Repeat Factor 2 (TRF2) in vitro.

METHODSFour different p53-GST (glutathione S-transferase) fusion proteins and GST were expressed in E. coli and purified through glutathione sepharose 4B beads. The human recombinant p53s included wild type p53 (1-393), N terminus-truncated form p53 2C (95-393), C terminus-truncated form p53 N5 (2-293) and single amino acid mutant p53 R175H (175 arginine to histidine). Purified p53-GST fusion proteins and GST were mixed with cellular protein extracts of human breast cancer cells MCF-7 in vitro by pull down. The molecular interaction between p53 and TRF2 were detected by Western blot.

RESULTSSDS-PAGE and Coomassie brilliant blue staining showed that the molecular weights of all purified proteins were as expected, with purities over 90%. Western blot of TRF2 indicated that both wild type p53 and p53 R175H could bind with TRF2 of MCF-7 cells in similar capacity, while GST alone failed to do so. The molecular interaction between p53 2C and TRF2 was enhanced. In contrast, the interaction between p53 N5 and TRF2 was significantly reduced.

CONCLUSIONSp53 can interact with TRF2 directly and specifically in vitro, with C terminus of p53 (293-393) being the binding region for their interaction. This C terminus-dependent interaction between p53 and TRF2 may be related to the cellular activities induced by telomere alterations.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; DNA-Binding Proteins ; metabolism ; Escherichia coli ; genetics ; metabolism ; Female ; Glutathione Transferase ; genetics ; metabolism ; Humans ; Protein Binding ; Recombinant Fusion Proteins ; isolation & purification ; metabolism ; Telomeric Repeat Binding Protein 2 ; metabolism ; Transformation, Genetic ; Tumor Suppressor Protein p53 ; genetics ; metabolism