Role of miR-214 in biomaterial transplantation therapy for osteonecrosis.

BACKGROUND: The effectiveness and availability of conservative therapies for osteonecrosis of the femoral head (ONFH) are limited. Transplantation of bone marrow mesenchymal stem cells (BMSCs) combined with Bio-Oss, which is a good bone scaffold biomaterial for cell proliferation and differentiation, is a new potential therapy. Of note, the expression of miRNAs was significantly modified in cells cultured with Bio-Oss, and MiR-214 was correlated positively with osteonecrosis. Furthermore, miR-214 was upregulated in cells exposed to Bio-Oss. OBJECTIVE: To investigate whether targeting miR-214 further improves the transplantation effect. METHODS: We treated BMSCs with agomiR-214 (a miR-214 agonist), antagomiR-214 (a miR-214 inhibitor), or vehicle, followed by their transplantation into ONFH model rats. RESULTS: Histological and histomorphometric data showed that bone formation was significantly increased in the experimental groups (Bio-Oss and BMSCs treated with antagomiR-214) compared with other groups. CONCLUSIONS: miR-214 participates in the inhibition of osteoblastic bone formation, and the inhibition of miR-214 to bone formation during transplantation therapy with Bio-Oss combined with BMSCs for ONFH.

Body Mass Index Changes in Relation to Male Reproductive Hormones: Longitudinal Results From a Community-Based Cohort Study.

The objective of the current study was to explore the relationship between longitudinal change in body mass index (BMI) and reproductive hormones in middle-aged and elderly Chinese men. A cohort study was conducted in a rural area of China. Local male residents aged 40-80 years were recruited at baseline in 2012 and were followed up in 2016. Information about weight, height, waist circumference, sex hormones, smoking status, and medical history were obtained. The change in BMI reported no significant relationship with the change in total testosterone (TT), calculated free testosterone (cFT), and bioavailable testosterone (BioT) in Pearson correlation analyses. When the change in BMI was divided into three groups-"great loss," "normal fluctuation," and "great gain"-TT, cFT and BioT had the highest increase (or the lowest decrease) in men with "normal fluctuation" in BMI compared with the other two groups. The advantage of maintaining a stable BMI was more evident for those who were overweight, non-smoking, and disease-free. There was a tendency of a continuous increase in cFT and BioT with BMI increase in smoking and diseased populations. Maintaining a stable BMI is associated with maintaining normal levels of reproductive hormones, especially in overweight, non-smoking, and healthy men aged over 40 years.

Kondo effect and superconductivity in niobium with iron impurities.

Kondo effect is an interesting phenomenon in quantum many-body physics. Niobium (Nb) is a conventional superconductor important for many superconducting device applications. It was long thought that the Kondo effect cannot be observed in Nb because the magnetic moment of a magnetic impurity, e.g. iron (Fe), would have been quenched in Nb. Here we report an observation of the Kondo effect in a Nb thin film structure. We found that by co-annealing Nb films with Fe in Argon gas at above 400 [Formula: see text]C for an hour, one can induce a Kondo effect in Nb. The Kondo effect is more pronounced at higher annealing temperature. The temperature dependence of the resistance suggests existence of remnant superconductivity at low temperatures even though the system never becomes superconducting. We find that the Hamann theory for the Kondo resistivity gives a satisfactory fitting to the result. The Hamann analysis gives a Kondo temperature for this Nb-Fe system at [Formula: see text] 16 K, well above the superconducting transition onset temperature 9 K of the starting Nb film, suggesting that the screening of the impurity spins is effective to allow Cooper pairs to form at low temperatures. We suggest that the mechanism by which the Fe impurities retain partially their magnetic moment is that they are located at the grain boundaries, not fully dissolved into the bcc lattice of Nb.

ELABELA improves endothelial cell function via the ELA-APJ axis by activating the PI3K/Akt signalling pathway in HUVECs and EA.hy926 cells.

Destruction of endothelial cells (ECs) function is involved in the structural and functional pathophysiological processes of preeclampsia (PE). Vascular endothelial injury may pre-exist for several years in women that develop PE and may pose increased risks for hypertension, coronary artery disease, and type-2 diabetes mellitus. Previous findings showed that Elabela (ELA), the endogenous ligand of the apelin (APJ) receptor expressed mainly on ECs, may play a protective role in early pregnancy and prevent PE. However, the exact functional role and molecular mechanisms of ELA are unclear. Here, we aimed to classify whether and how ELA improves EC function via the ELA-APJ axis. Two human umbilical vein endothelial cell (HUVEC) lines, namely HUVECs and EA.hy926, were treated with ELA, and then their cellular activities were studied by performing CCK-8 tests, scratch-wound analysis, and tube-formation assays. Doses of ELA exceeding 0.01 µmol/L markedly improved the cell viability, migration, and tube formation ability of HUVECs and EA.hy926 cells. Western blot analysis indicated that the above effects caused by ELA were related to upregulation of the APJ receptor and activation of PI3K/Akt signalling. Further verification tests were performed using the PI3K inhibitor wortmannin, and the results illustrated that inhibiting PI3K/Akt signalling blocked the positive effects of ELA on EC function and APJ receptor expression. Taken together, our findings indicate that ELA may alter EC function via the ELA-APJ axis and PI3K/Akt signalling and that ELA shows promise for use in endothelial dysfunction therapy for preventing and treating PE.

Hypoxia regulates angeogenic-osteogenic coupling process via up-regulating IL-6 and IL-8 in human osteoblastic cells through hypoxia-inducible factor-1α pathway.

Inappropriate angiogenesis and osteogenesis are considered as the crucial factors of osteoporotic fracture. Hypoxia is a primary driving force for regulating the angiogenic-osteogenic coupling process. Our recent results indicated that hypoxia could improve angiogenesis as well as differentiation and activity of osteoblastic cells via up-regulating VEGF through HIF-1α pathway. Here we demonstrated that in human osteoblastic MG-63, U2-OS and Saos-2 cells, besides VEGF, the other two pro-angiogenic factors IL-6 and IL-8 were also up-regulated by hypoxia and CoCl2 (a mimic of hypoxia). Mechanism studies indicated overexpression of HIF-1α (generated from transfection with a plasmid encoding sense HIF-1α) markedly increased the levels of IL-6 and IL-8 in osteoblastic cells. Furthermore, a luciferase reporter assay was performed using the reporter vector containing the IL-6 or IL-8 promoter sequence to illustrate observably increased activity of hypoxia-induced IL-6 and IL-8 promoter caused by overexpression of HIF-1α. Additionally, chromatin immune-precipitation analysis showed hypoxia increased the DNA binding ability of HIF-1α to IL-6 or IL-8 promoter. Analysis in vitro by MTT test and Boyden chamber assay showed exogenous IL-6 and IL-8 (a relatively short period of treatment with recombinant IL-6 or IL-8 equivalent to the autocrine levels) could significantly promote the proliferation of human osteoblastic, endothelial and monocytic cells, as well as the migration of human endothelial cells. Taken together, these results indicate that IL-6 and IL-8 in osteoblastic cells may also contribute to the angiogenic-osteogenic coupling process via HIF-1α pathway. Besides VEGF, IL-6- or IL-8-targeted adjunctive therapy maybe a new strategy to improve the treatment of osteoporosis.

Seminal plasma miR-192a: a biomarker predicting successful resolution of nonobstructive azoospermia following varicocele repair.

This study was performed to investigate a potential marker for the presence of spermatozoa in the ejaculate following varicocelectomy in Chinese men with nonobstructive azoospermia and varicoceles. The micro-RNA (miR)-192a levels in seminal plasma and testicular tissue were evaluated by quantitative real-time polymerase chain reaction from 60 men with nonobstructive azoospermia and varicoceles (Group A: 27 men with spermatozoa found in the ejaculate after surgery; Group B: 33 men without spermatozoa found in the ejaculate after surgery) and 30 controls. The seminal plasma and testicular tissue miR-192a levels were higher in Group B than in Group A and the controls (P < 0.001), and there was no significant difference between Group A and the controls (P > 0.05). Apoptosis and proliferation assays with miR mimics and inhibitors showed that miR-192a induced GC-2 cell apoptosis through the activation of Caspase-3 protein. Thus, seminal plasma miR-192a appears to be a potential marker for successfully indicating spermatozoa in the ejaculate following microsurgical varicocelectomy in men with nonobstructive azoospermia and varicoceles. Seminal plasma miR-192a may be a useful clinical marker for prescreening to determine which patients with nonobstructive azoospermia and varicoceles would benefit from varicocelectomy.

[The expression of Calbindin and Parvalbumin in auditory pathway of kit gene mutated C57BL/6J mouse].

OBJECTIVE: To observe the expressions of Calbindin(CB) and Parvalbumin (PV), the two calcium-binding protein, in auditory pathway in mice of wild type C57BL/6J and kit⁺/kitW⁻ ²Bao, a kit gene mutant. METHODS: Six mutated kit gene kit⁺/kitW⁻ ²Bao mice and 6 wild type C57BL/6J (B6) mice were anaesthetized i. p. with chloral hydrate. After the mice were fixed by heart perfusion, the brains were removed and coronal sections were cut with a freezing microtome. RESULTS: We found that wild type mice had significant expressions of PV on ventral cochlear nucleus, anterior part (AVCN), ventral cochlear nucleus, posterior part (PVCN), inferior colliculus (IC) and auditory cortex (AC). CB was expressed in wild type mice on PVCN and nucleus of the trapezoid body (Tz). The mutant of kit gene induced the less expression of PV on PVCN, IC and AC (P < 0.01), but increased the expression of Tz (P < 0.01). CB could not be observed on PVCN in mutant mice, and the expression of AC was increased( P < 0.01). CONCLUSION: CB and PV has differential expression level in auditory pathway. Since mutated kit gene can affect expression of PV on PVCN, IC, Tz and AC, as well as CB on PVCN and AC, it suggests that the mutation of kit gene can affect the advanced function of central nervous system in auditory pathway.

The relationship between physical activity and aging symptoms among community-dwelling men aged 40-70 years in Shanghai, China.

BACKGROUND: Research on the relationship between physical activity and aging symptoms among men is limited in China. METHODS: Eight hundred nineteen participants aged 40 to 70 (mean age = 58.25) were administered the International Physical Activity Questionnaire and Aging Males' Symptoms scale (AMS) in face-to-face interviews between 2009 and 2010. Ordinal logistic regression and generalized linear models were used to examine severity of aging symptoms. AMS total score and subscale scores (psychological, somatic, and sexual health) were examined according to physical activity level. RESULTS: Participants who reported higher levels of physical activity were significantly less likely to develop aging symptoms compared with those who reported lower levels of physical activity, both with (odds ratio [OR] = 0.78, 95% confidence interval [CI]: 0.64 to 0.95) and without (OR = 0.70, 95% CI: 0.56 to 0.87) adjustment for age, body mass index (BMI), smoking, alcohol consumption, and chronic disease. In addition, physical activity was negatively correlated with AMS total score (P < .01) and scores for somatic (P < .01) and sexual (P = .04) health. After covariate adjustment, the negative correlation between physical activity and all AMS scores remained significant (P < .01). CONCLUSION: Physical activity is negatively correlated with aging symptoms among middle-aged men in Shanghai.

Relationship between age and prostate size.

In a community-based study, the relationship between age and human prostate size was investigated in a population of men between the ages of 40 and 70 years to determine the normal prostate increase curve equation. One thousand male volunteers were randomly recruited from the Shanghai community, and the length, width, height, volume of the transition zone (TZ) and the whole prostates were measured by transrectal ultrasound (TRUS). Each volunteer was evaluated by the International Prostate Symptom Score (IPSS). Among those who completed the examination, the mean prostate parameters were all positively associated with increased age. There were statistically significant differences between each age group (P<0.05). The mean transition zone volume (TZV) had a higher increase rate with age than the mean total prostate volume (TPV), indicating that the enlargement of the TZ contributed the most to the increase in TPV. While all prostate parameters were positively correlated with the IPSS, the strongest correlation was associated with the TZ length (TZL) and TZV. The growth curve equations for prostate width, height and length were also positively associated with increasing age.

Application of pyrosequencing technique for improved detection of K-Ras mutation in formalin-fixed and paraffin-embedded prostate carcinoma tissues in Chinese patients.

BACKGROUND: The purpose of this study was to comparatively evaluate the efficacy of pyrosequencing (PS) and Sanger sequencing (SS) methods for detecting K-Ras codon 12 and 13 mutations in formalin-fixed paraffin-embedded (FFPE) prostate cancer (PCa) samples from Chinese patients. METHODS: The cancer cell lines, including the LS174T G12D mutant (GGT to GAT) and the COLO320 wild-type, were tested to determine the limitation of detection and reproducibility of the PS method. In addition, 101 PCa patient samples, 13 benign prostatic hyperplasia (BPH) and 12 normal adjacent tissue samples, were assayed by PS and SS to evaluate their detection abilities for K-Ras mutations in codons 12 and 13. RESULTS: The PS assay was able to reproducibly detect 5% mutant alleles and had an intra-assay variability of 4.21% and inter-assay variability of 11.37%. The PS assay detected a higher number of K-Ras mutations in PCa samples than the SS assay (8.91% vs. 3.96%). Correlation and stratification analyses of the PCa samples and K-Ras mutation status revealed no associations between age, serum prostate specific antigen (PSA), depth of invasion (pT category), or Gleason score. CONCLUSIONS: We demonstrated that the PS method detected more K-Ras mutations in codons 12 and 13 of FFPE prostate cancer samples from Chinese patients than the traditional SS method. In addition, the K-Ras mutation was more frequent in Chinese population than in Western populations but was similar to that of other Eastern populations, suggesting that these K-Ras mutations may contribute to the pathogenesis of prostate carcinoma in Asian patients.

[Lower urinary tract symptoms and prostatic growth pattern among old and middle-aged males in Shanghai community].

OBJECTIVE: To investigate the prevalence of lower urinary tract symptoms (LUTS) and the age-related growth pattern of the prostate among 40 -70 year-old males in Shanghai community. METHODS: Using cluster and stratified random sampling and IPSS, we investigated the prevalence of LUTS among 1000 males aged 40 -70 years in the general population of Shanghai from November 2009 to June 2010. We measured the transverse, anteroposterior and vertical diameters of the prostate and its transition zone in each volunteer by transrectal ultrasonography and established the equation for the age-related growth pattern of the prostate. RESULTS: In the 40 to 49-, 50 to 59- and 60 to 70-year groups, the incidence rates of moderate and severe LUTS (IPSS > or = 8) were 10.0%, 15.0% and 28.7%, respectively. The length, width, height and volume of the prostate and its transition zone were positively corrected with age (P < 0.05). The prostatic growth pattern equations based on the parameters of the transverse, anteroposterior and vertical diameters were Y = 1.6 x 10(-5)X3-0.002 1X2 + 0.074 6X + 0.677 2, Y = -2.4 x 10(-5)X3 + 0.003 3X2-0.1312X + 1.269, and Y = 1.6 x 10(-5)X3-0.001 8X2 + 0.073X- 0.690 9, respectively. The transverse and anteroposterior diameters of the prostate grew at a relatively similar rate, while the transverse diameter grew obviously faster than the vertical diameter before 60 years old, but the latter significantly increased and even exceeded the former after 60 years old. CONCLUSION: The prevalence of LUTS among old and middle-aged males in Shanghai community is similar to that recently reported at home and abroad. The transverse and anteroposterior diameters of the prostate grow at a relatively similar rate, but the vertical diameter increases faster after 60 years old.

[Benefits of placing intra-aortic vacant guide wire on interventional treatment of aorto-ostial lesions].

OBJECTIVE: To evaluate the potential benefits of placing intra-aortic vacant guide wire on interventional treatment of aorto-ostial lesions. METHODS: A total of 86 consecutive patients underwent percutaneous coronary interventions (PCI) for aorto-ostial lesions were randomly divided into conventional treatment group (group A) and intra-aortic vacant guide wire group (group B). Standard PCI techniques were applied in group A and an intra-aortic vacant guide wire was placed outside the guiding catheter before the guiding catheter into the target coronary artery in group B on basis of standard PCI techniques. The number of guiding catheter, guide wire, X-ray exposure time, total PCI time, the incidence of pressure drop and malignant arrhythmia, contrast agent dose and the number of failure cases were compared between the two groups. RESULTS: The number of used guide wire was similar between the groups (2.0 ± 1.1 vs. 2.2 ± 0.4, P > 0.05) and the number of guiding catheter used was significantly more in group A than in group B (2.3 ± 1.1 vs. 1.3 ± 0.5, P < 0.01). The X-ray exposure time (18.8 min ± 6.9 min vs. 14.2 min ± 5.7 min, P < 0.01) was significantly less in groups B and the total PCI time (31.2 min ± 8.1 min vs. 20.1 min ± 4.5 min, P < 0.01) and the amount of contrast agent (193.5 ml ± 25.4 ml vs. 130.6 ml ± 32.8 ml, P < 0.01) and the frequency of pressure drop (19 cases vs. 2 cases, P < 0.01) were all significantly higher in group A than in group B. Incidence of malignant ventricular arrhythmia (8 vs. 0) and procedure failure (6 vs. 0) was also higher in group A compared to group B. CONCLUSION: The method of using Intra-aortic vacant guide wire is associated with reduced risk and improved success rate of PCI for aorto-ostial lesions.

[Pulmonary manifestations of Wegener's granulomatosis].

OBJECTIVE: To study the clinical features of pulmonary Wegener's granulomatosis. METHODS: The data of 20 patients with Wegener's granulomatosis diagnosed during the last 20 years were retrospectively analyzed. RESULTS: (1) Twelve patients (60%) with Wegener's granulomatosis presented with symptoms of respiratory system including cough, purulent sputum, bloody sputum, hemoptysis or dyspnea. In 5 of the 12 patients, the pulmonary symptoms presented as the initial manifestations. (2) Chest radiography demonstrated that Wegener's granulomatosis could involve multiple lung fields, either unilateral or bilateral in the lung lobes, but bilateral lung involvement (8/12) was more common. The lesions included nodules, masses, thick or thin wall cavities, or infiltrates. (3) Multiple biopsies for histologic study were done in 14 patients. CONCLUSIONS: Wegener's granulomatosis has different radiographic manifestations and is difficult to differentiate from pulmonary abscess, tuberculosis, pneumonia or lung cancer. Repeated and multiple biopsies are essential to confirm the diagnosis.