Objective To show the international protocols for blood pressure monitoring based on a real example. Methods The assessment process of international protocol that can be released by Working Group on Blood Pressure Monitoring of European Society of Hypertension was evaluated. Results 33 participants were selected, which all indexes in evaluation stage one and stage two of the indicators were detected through. The 95% consistency interval in difference between tested device and reference monitor was 10.65～-12.67 mmHg for systolic BP and 13.68～-14.03 mmHg for diastolic BP, and there were 7.1% (7/99) and 6.1% (6/99) of valid points out of the 95% consistency interval. Conclusion The measured automatic blood pressure in the normal environment, measuring accuracy and the standard with the control of mercury -type sphygmomanometer is coincident, so it can be recommended for home application.

Objective To compare blood pressures results measured by automated sphygmomanometer and standard mercury sphygmomanometer,and to investigate the application of measurements consistency evaluation method in accurate measurement of automated sphygmomanometer.Methods Intraclass correlation coefficient was used to estimate the reliability of repeated measurements,and Bland -Altman method was adopted to evaluate the consistency between automated sphygmomanometer and standard mercury sphygmomanometer.Meanwhile,the results were compared with protocol of European Society of Hypertension.Results The tested automated sphygmomanometer did not adapt to the criteria of European Society of Hypertension.The intraclass correlation coefficient of mercury sphygmomanometer was 0.937 for systolic blood pressure,0.849 for diastolic blood pressure.The intraclass correlation coefficient of tested sphygmomanometer was 0.944 for systolic blood pressure,0.929 for diastolic blood pressure.The 95% consistency interval was(-10.20 to 16.94)mmHg for systolic blood pressure and(-6.25 to 11.69)mmHg for diastolic blood pressure.Conclusion Normally,Bland-Altman method has the same judgment result with protocol of European Society of Hypertension.

BACKGROUND:Stem celltumorigenicity is a practical issue concerning stability in the clinical application of stem cells. Therefore, it is particularly important to clear whether stem cells have tumorigenic ability or not. Nude mice occupy an increasingly important position in oncology, immunology, and safety evaluation of drugs and biological products.
OBJECTIVE:To observe the tumorigenicity of neural stem cells and mesenchymal stem cells in Balb/c nude immunodeficient mice.
METHODS:Balb/c nude mice were randomly divided into control group, negative group, positive group, neural stem cellgroup and mesenchymal stem cellgroup. HepG-2 cells, RPE cells, passage 4 neural stem cells and mesenchymal stem cells were injected subcutaneously into nude mice from different groups, respectively. After 12 weeks of injection, anatomical observation was performed to detect the tumor formation in the transplantation site. Meanwhile, soft agar colony formation assay was applied to investigate neural stem celland mesenchymal stem cellclone in vitro.
RESULTS AND CONCLUSION:After 12 weeks of injection, the tumorigenicity study results showed that no tumor developed in the transplantation site in the control group, negative group, neural stem cellgroup and mesenchymal stem cellgroup. Histopathologic examinations also showed no abnormality in these groups. Soft agar colony formation assay showed in soft agar resistance medium, neural stem cells and mesenchymal stem cells did not exhibit clone ability. These findings indicate that neural stem cells and mesenchymal stem cells undergoing short-term passages have no tumorigenic growth.

BACKGROUND:Stem cells expansion technology in vitro is affected by the microenvironment of the culture system. So, to find an effective method is particularly important to promote celladherent and growth. OBJECTIVE:To compare the effects of different culture media on cellexpansion. METHODS:Human umbilical cord mesenchymal stem cells at a density of 5.0×104 cells/cm2 were inoculated into Corning? polystyrene culture dishes coated with or without poly-L-ornithine and Corning? cellBIND culture dishes. celladhesion and proliferation were observed, and expressions of celladhesion proteins and cellmarkers were detected. RESULTS AND CONCLUSION:celladhesion was promoted when cells were cultured in Corning? cellBIND? Surface medium coated with poly-L-ornithine for 24 hours, and the cultured cells grew at the logarithmic phase. The cellproliferation was also enhanced, and the cells expressed celladhesion protein but not the cellmarkers of CD73, CD90, CD105. Corning? cellBIND? Surface medium was superior to Corning? polystyrene culture medium in the improvement of celladhesion and proliferation. Additional y, both of these two media showed no influence on cellphenotype. These findings indicate that Corning? cellBIND? Surface medium can promote celladhesion and proliferation, but shows no effects on cyclin and cellphenotype. This medium coated with poly-L-ornithine can further accelerate celladhesion and proliferation, and stably express cellphenotype of human umbilical cord mesenchymal stem cells.

Objective To explore the effect of excessive fluoride on expression of mRNA and protein of Wnt3a and β-catenin in rats' osteoblasts and its correlation with pathogenic mechanism of fluorosis.Methods Thirty-six healthy SD rats,weighting 100-120 g and according to body mass,were randomly divided into three groups(twelve in each group).The rats of control were fed wich tap water(fluoride ＜ 1 mg/L) and the experimental rats were exposed to NaF(low-fluoride group:5 mg/L,high-fluoride group:50 mg/L) added to the drinking water to establish the chronic fluorosis model.After fed for eight morth,all rats were killed and metaphysic of femoral was collected.Rat dental fluorosis was observed and bone fluorine was detected by ashing-fluorin ion selective electrode method.The content of bone alkaline phosphatase (BALP) and tartrate-resistant acid phosphatase 5b(TRACP 5b) in rats' serum was detected by enzyme-linked immunosorbent assay (ELISA).The morphologic changes of the bone were observed by microscopy.The expression of mRNA and protein of Wnt3a and β-catenin in osteoblasts of rats was analyzed with gray scale by hybridization in situ and immunohistochemistry methods,respectively.Results Detection rate of dental fluorosis,fluoride contents of urine and bone were significantly increased [control group:0.0％,(1.26 + 0.17)mg/L,(305.58 ± 91.26)mg/kg; low-fluoride group:66.7％,(2.06 ± 0.64)mg/L,(632.33 ±123.21)mg/kg; high-fluoride group:91.7％,(7.69 ± 1.96)mg/L,(1088.75 ± 156.16) mg/kg] in the rats treated with fluoride,the difference between groups was statistically significant(χ2 =21.6; F =36.57,467.02; all P ＜0.05).The contents of BALP and TRACP-5b in rats' serum were significantly different between groups(F =89.57,7.68; all P ＜ 0.05).Compared with control group[(16.24 + 1.57)U/L],the contents of BALP in rats' serum of the low-fluoride and high-fluoride groups[(31.47 ± 5.30) and (54.61 ± 2.27)U/L] were increased gradually(all P ＜0.05).Compared with the low-fluoride group,the value in the high-fluoride group decreased significantly (P ＜ 0.05).The contents of TRACP-5b in rats' serum of low-fluoride group[(3.45 ± 1.85)U/L] were elevated significantly(all P ＜ 0.05) compared with the control group[(1.26 ± 0.23)U/L] and the high-fluoride group[(2.74 ± 1.85)U/L].The bone cortices were thickened and the bone trabecula was broadened,arranged closely together in chronic fluorosis rats with significant difference compared with the control group.In the low-fluoride and high-fluoride groups,the expression levels of Wnt3a and β-catenin mRNA (low-fluoride group:132.87 ± 5.72 and 132.57 ± 9.56; highfluoride group:135.60 ± 6.64 and 137.87 ± 9.16) were markedly elevated with significant difference,respectively (F =12.47,5.96; all P ＜ 0.05) compared with those in control groups(119.86 ± 5.04 and 120.58 ± 7.84) by hybridization in situ(P ＜ 0.05),but there was no statistical significance (P ＞ 0.05) of the level of Wnt3a and β-catenin mRNA between low-fluoride and high-fluoride groups.In the low-fluoride and high-fluoride groups,the protein expression of Wnt3a and β-catenin (low-fluoride group:137.50 ± 4.32 and 140.85 + 3.54; high-fluoride group:142.65 ± 11.84 and 152.52 ± 4.64) were markedly elevated with significant difference,respectively (F =10.07,53.82; all P ＜ 0.05) compared with those in control group (124.01 ± 2.63 and 126.75 ± 4.65) by immunohistochemistry(all P＜ 0.05),Wnt3a protein production in the low-fluoride group was increased without statistical significance compared with the high-fluoride group (P ＞ 0.05).But the protein production of β-catenin in the lowfluoride group was elevated with significant difference compared with the high-fluoride group(P ＜ 0.05).The mRNA and protein production of Wnt3a were positively correlated with the mRNA and protein production of β-catenin (r =0.731,0.658; all P ＜ 0.05).Conclusions Rat bone tissue lesions caused by excessive fluoride may be associated with an increased expression of Wnt3a and β-catenin mRNA and protein in osteoblasts.In chronic fluorosis,fluoride stimulates the overexpression of Wnt3a and β-catenin in the Wnt signal transduction pathway,enhances bone osteogenesis and causes skeletal fluorosis.

AIM:To examine the difference of vascular remodeling between aorta and small artery in spontaneous hypertensive rats (SHR) and control rats.METHODS:Male SHR (20-week-old) were used as experiment group,and age matched male Wistar-Kyoto (WKY) rats were used as control group.The systolic blood pressure and body weight were measured once a week.At 43 weeks old,the rats were anaesthetized,blood samples were collected,and thoracic aorta and mesenteric small artery tissue were harvested.The morphological changes of the arterial tissue were observed with HE staining.The collagen and elastine fibers were detected by the Sirius red-Victoria blue staining.The protein expression of type Ⅰ and Ⅲ collagens were analyzed by confocal laser-scanning microscopy and Western blot.The changes of the vascular ultrastructure were imaged by transmission electron microscopy.The expression of proliferating cell nuclear antigen (PCNA) and the cell apoptosis in the arterial wall were examined by immunohistochemical method and TdT-mediated dUTP nick and labeling (TUNEL) detection.RESULTS:The inner diameter (ID) and luminal cross-sectional area (LCSA) of mesenteric small artery were decreased,whereas ratio of wall thickness (WT) to ID (WT/ID) and ratio of wall cross-sectional area (WCSA) to LCSA (WCSA/LCSA) were increased.Meanwhile,adventitia fibroblast migrated to the nedia,with overload collagens,especially collagen Ⅲ.Proliferation index (PI) and apoptotic index (AI) of the mesenteric small artery wall cells were increased.The ID,LCSA,WT/ID and WCSA/LCSA of the aorta were increased.Moreover,the vascular smooth muscle cells (VSMCs) showed hypertrophy and hyperplasia,with overload collagens.The PI and AI of the aortic wall cells were increased.CONCLUSION:The difference of vascular remodeling between the aorta and small artery is significant.The small artery mainly appears hyperplasia of matrix,especially the adventitial collagen Ⅲ.Meanwhile,the cell apoptosis in the small artery wall is increased.The aorta mainly appears hyperplasia and hypertrophy of media VSMCs.

OBJECTIVE: To summarize the subtle anatomical structures of the normal nasal bone in multi-slice spiral CT (MSCT) image through the observation of the three-dimensional images. METHODS: One hundred and twenty volunteers who had no nasal trauma and disease history were collected. The nasal was scanned using MSCT. Raw data was reconstructed into bone window images (slice thickness 0.6 mm, slice interval 0.5 mm), and then the images were imported into Syngo Imaging XS software to reconstruct three-dimensional images and to summarize the nasal bone's subtle anatomical structures. RESULTS: The subtle anatomy of normal nasal bone generally included four seams, two holes and one edge. The four seams were left and right nasal-maxillary suture, nasal-frontal seam, and internasal suture. The two holes were left and right nasal aperture. The edge of the nasal was the lower edge of the nasal bone. In addition, there was suture bone in internasal suture in some normal nose. The nasal aperture mostly was hole-like, but some nasal apertures were line shape. The nasal edge can be divided into flat type, wave-shaped type, inverted spike type, hook-shaped type and others. CONCLUSION The anatomy diversity and individual differences in nasal bone are large. MSCT and three-dimensional image reconstruction techniques, combined with the history of trauma could distinguish between the normal anatomy and fractures.
Fractures, Bone/diagnosis* ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; Maxilla ; Nasal Bone/diagnostic imaging* ; Software ; Tomography, Spiral Computed ; Tomography, X-Ray Computed

OBJECTIVE: To realize the automated assessment of the levels of epiphysis of distal radius and ulna by support vector machine (SVM). METHODS: The X-ray films of the left wrist joints were taken from 140 teenagers aged from 11 to 19 years old as training samples. The levels of epiphysis of distal radius and ulna were divided into five developmental levels. Each level contained 28 samples. Another 35 cases were selected as independent verifying samples. SVM classification models of the five developmental levels of epiphysis of distal radius and ulna were established. The internal cross validation was made by leave one out cross validation (LOOCV), while the external validation was made by histogram of oriented gradient (HOG), and then the accuracy (PA) of testing results was calculated, respectively. RESULTS: The PA of SVM, LOOCV and HOG of distal radius epiphyseal level were 100%, 78.6%, and 82.8%, respectively; whereas the PA of SVM, LOOCV and HOG of distal ulna epiphyseal level were 100.0%, 80.0% and 88.6%, respectively. CONCLUSION The SVM-based automatic models of the growth stage of distal ra- dius and ulna appear to have certain feasibility, and may provide a foundation for software development of bone age assessment by forensic medicine.
Adolescent ; Bone Development/physiology* ; Child ; Epiphyses/growth & development* ; Female ; Humans ; Image Processing, Computer-Assisted/methods* ; Male ; Radius/growth & development* ; Support Vector Machine ; Ulna/growth & development* ; Wrist/growth & development* ; Wrist Joint/growth & development* ; Young Adult

OBJECTIVETo investigate the changes of mRNA and protein expression of CaN in the bone of rats with chronic fluorosis, and the mechanism of skeletal fluorosis.

METHODSThirty-six SD rats were divided into three groups (12 in each group, half male and half female selected according to body weight): control, low-dose and high-dose fluorosis groups. Controls were fed tap water (NaF < 0.5 mg/L), experimental animals in the low- or high-dose groups were fed water containing NaF of 5.0 and 50.0 mg/L, respectively. The rats were sacrificed after 6 months of treatment with fluoride. The serum was kept for testing bone metabolic marker bone gla protein (BGP) by enzyme-linked immunosorbent assay (ELISA), the protein and mRNA levels of CaN in distal femur of the rats with chronic flurosis were assessed by immunohistochemistry and in-situ hybridization.

RESULTSThe levels of BGP (1.99 ± 0.62, 2.38 ± 0.16)µg/L in the low- or high-dose fluorosis groups were higher than that in the control group (0.15 ± 0.03) µg/L; and the high fluorosis group showed higher level than the low fluorosis group (all P < 0.05). Compared to the control group (131.11 ± 1.95, 111.82 ± 2.39), the protein and mRNA levels of CaN were higher in the low- or high-dose fluorosis groups (142.69 ± 1.17, 157.54 ± 1.88 and 121.28 ± 3.27, 134.63 ± 3.19, respectively), and the high fluorosis group showed higher levels than the low fluorosis group (all P < 0.05).

CONCLUSIONSBGP content could be used as a bone metabolic index in endemic fluorosis disease. Fluoride might up-regulate the mRNA and protein expression of CaN, and the changes in CaN level may be involved in the increase of the bone turnover and could be one of the pathogenetic factors in fluorosis.

Animals ; Bone and Bones ; metabolism ; Calcineurin ; genetics ; metabolism ; Female ; Fluoride Poisoning ; metabolism ; pathology ; Fluorides ; metabolism ; urine ; Fluorosis, Dental ; metabolism ; pathology ; Male ; Osteoblasts ; metabolism ; Osteocalcin ; blood ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sodium Fluoride ; poisoning

Tumor immune checkpoint therapy is a clinical treatment strategy developed based on the new principle of the inhibition of negative immune regulation. In this article, the tumor immune checkpoint therapy and the drug delivery strategies were reviewed, mainly including immunity and tumor therapy, tumor immune checkpoint therapy and its mechanism of action, clinical application of tumor immune checkpoint therapy and therapeutic drugs, immune resistance of programmed cell death protein 1 (PD1)/programmed cell death ligand 1 (PDL1) treatment and countermeasures, drug delivery strategies for tumor immune checkpoint therapeutic agents, etc. As a revolutionary new immunotherapy strategy, tumor immune checkpoint therapy has shown obvious superior therapeutic efficacy in a variety types of tumor. However, tumor immune checkpoint therapy is also faced with a big challenge, namely, immunotherapy resistance. With the discovery of new mechanism, the continuous development of new therapeutic drugs and delivery strategies, tumor immune checkpoint therapy is expected to further improve the clinical efficacy of tumor.