G/ ß- TCP composite scaffold material promotes osteogenic differentiation of bone marrow mesenchymal stem cells.

To explore self-made graphene/ß Graphene (G)/ß- tricalcium phosphate, G/ß- The effect of TCP composite scaffold material on osteogenic differentiation of BMSC. Preparation of G/ß- TCP composite material was used to investigate the effect of composite material on bone marrow mesenchymal stem cell ossification/ß- TCP material was used to treat primary BMSCs of rats. Cell morphology changes were observed under scanning electron microscopy, cell cycle and proliferation were detected by flow cytometry, and gene expression of chondrogenic genes Fibronectin, collagen I, collagen II, ICAM, and VCAM was detected by q-PCR. In addition, using osteogenic induction medium and G/ß- TCP composite materials were co treated with BMSCs, and ALP and alizarin red staining were used to observe the effect of the materials on osteogenic differentiation. q-PCR was used to detect the gene expression of osteogenic related genes Runx2, OCN, and OPN. G/ ß- After the TCP composite was co cultured with BMSC, the proportion of G0/G1 phase of BMSC cells was significantly increased, the cell proliferation ability was enhanced, and the gene expression of fibronectin, collagen I, collagen II, ICAM, and VCAM were significantly increased. The ALP staining results indicate that BMSC in G/ß- After treatment with TCP composite material, significant enhancement of osteogenic ability was observed at 7,14 and 21 days. In addition, BMSC in G/ß- A significant increase in calcium deposition was observed at 7,14 and 21 days after treatment with TCP composite materials. The effect of different time points on the expression of osteogenic related genes varies. At 7 and 14 days, the expression of RUNX2 was significantly reduced compared to the control, but significantly increased at 21 days; OCN significantly increased on the 21st day; OPN significantly increased at 14 days. G/ß- TCP materials significantly promote the osteogenic differentiation of BMSCs.

Bone marrow mesenchymal stem cell-derived exosomal microRNA-382 promotes osteogenesis in osteoblast via regulation of SLIT2.

BACKGROUND: Osteoporosis (OP) is a systemic skeletal disorder with increased bone fragility. Human bone marrow mesenchymal stem cells (hBMSCs) have multi-lineage differentiation ability, which may play important roles in osteoporosis. In this study, we aim to investigate the role of hBMSC-derived miR-382 in osteogenic differentiation. METHODS: The miRNA and mRNA expressions in peripheral blood monocytes between persons with high or low bone mineral density (BMD) were compared. Then we collected the hBMSC-secreted sEV and examined the dominant components. The over-expression of the miR-382 in MG63 cell and its progression of osteogenic differentiation were investigated by qRT-PCR, western blot and alizarin red staining. The interaction between miR-382 and SLIT2 was confirmed by dual-luciferase assay. The role of SLIT2 was also confirmed through up-regulation in MG63 cell, and the osteogenic differentiation-associated gene and protein were tested. RESULTS: According to bioinformatic analysis, a series of differential expressed genes between persons with high or low BMD were compared. After internalization of hBMSC-sEV in MG63 cells, we observed that the ability of osteogenic differentiation was significantly enhanced. Similarly, after up-regulation of miR-382 in MG63 cells, osteogenic differentiation was also promoted. According to the dual-luciferase assay, the targeting function of miR-382 in SLIT2 was demonstrated. Moreover, the benefits of hBMSC-sEV in osteogenesis were abrogated through up-regulation of SLIT2. CONCLUSION: Our study provided evidence that miR-382-contained hBMSC-sEV held great promise in osteogenic differentiation in MG63 cells after internalization by targeting SLIT2, which can be served as molecular targets to develop effective therapy.

Serum calcium concentration as an indicator of intervertebral disk degeneration prognosis.

This study aims to investigate the relationship between serum macro- and trace element contents and the degree of disk degeneration in patients with intervertebral disk herniation (IDH). This study was carried out on 69 subjects (30 women and 39 men) diagnosed with IDH. Blood samples of the subjects were collected, and serum concentrations of the elements that include macroelements, such as calcium, phosphorus, potassium, sodium, and magnesium, and trace elements, such as zinc, iron, copper, and selenium, were determined using inductively coupled plasma-atomic emission spectrometry. Magnetic resonance imaging (MRI) examination of the entire lumbar region of the vertebral column was conducted using a 1.5-T MRI scanner. The degree of disk degeneration was classified into three categories. Correlation analysis between the degree of disk degeneration and the serum element was performed using SPSS 16.0. In the correlation analysis between the degree of disk degeneration and the element contents, only calcium was found to be negatively correlated with the degree of disk degeneration (r = -0.332, P < 0.01). Comparison results between male and female groups showed no significant difference in the element content and in the degree of disk degeneration (P > 0.05). Moreover, the serum calcium content showed a significant correlation with the degree of disk degeneration, suggesting that the serum calcium concentration can be used as an indicator of intervertebral disk degeneration prognosis.

The ratio of sTfR/ferritin is associated with the expression level of TfR in rat bone marrow cells after endurance exercise.

Currently, it is unclear which index of haematological parameters could be used to most easily monitor iron deficiency during endurance training. To address this question, 16 male Wistar rats were randomly assigned to two groups: a sedentary group (n = 8) and an exercised group (n = 8). Initially, animals in the exercise group started running on a treadmill at a rate of 30 m/min, on a 0% grade, for 1 min/session. Running time was gradually increased by 2 min/day. The training plan was one session per day during the initial 2 weeks and two sessions per day during the third to ninth week. At the end of the 9-week experiment, we analysed the blood of the experimental animals for haemoglobin levels, erythrocyte numbers, haematocrit, serum iron levels, total iron binding capacity, transferrin saturation, serum ferritin levels and soluble transferrin receptor (sTfR) levels, and we calculated the ratio of sTfR/ferritin. Erythrocyte numbers, haemoglobin levels and haematocrit values were decreased after 9 weeks of exercise, but sTfR and sTfR/ferritin values were increased (P < 0.01 or P < 0.05). The training regime significantly increased TfR mRNA levels in the bone marrow cells of the exercised rats compared with the sedentary group (1.8 ± 0.5 vs. 1.1 ± 0.2, P < 0.01). These results revealed a significant correlation between TfR levels in the bone marrow cells and the ratio of sTfR/ferritin (r = 0.517; P < 0.01) and sTfR levels (r = 0.206; P < 0.05) in sedentary and exercised rats. In conclusion, we show that sTfR indices and the ratio of sTfR/ferritin could be useful indicators for monitoring iron deficiency during endurance training.

Endurance exercise is a leptin signaling mimetic in hypothalamus of Wistar rats.

BACKGROUND: Endurance exercise is known to promote a substantial effect on the energy balance in rats and humans. However, little is known about the exact mechanisms for the appetite-suppressive effects of endurance exercise. We hypothesized that endurance training might activate signaling cascades in the hypothalamus known to be involved in leptin signaling. METHODS: 16 male Wistar rats were randomly assigned to two groups: sedentary (n = 8) and exercise groups (n = 8). Animals in the exercise group started treadmill running at 30 m/min, 0% grade, for 1 min/bout. Running time was gradually increased by 2 min/bout every day. The training plan was one bout per day during initial two weeks, and two bouts per day during 3rd-9th week. At the end of nine-week experiment, blood was analyzed for low-density lipoprotein cholesterol (LDL-C), triglyceride (TG), total cholesterol (TC), free fatty acid (FFA), interleukin (IL)-6, and leptin in both groups. Activations of janus kinase 2-signaling transducer and activator of transcription 3 (JAK2-STAT3), protein kinase B (Akt), extracellular regulated kninase (ERKs), and suppressor of cytokine signaling 3 (SOCS3) in hypothalamus were measured in the end of nine weeks of exercise protocol. RESULTS: Nine-week endurance exercise induced lower concentrations of LDL-C, TG, TC, FFA, and leptin in rats (P < 0.05 or P < 0.01). Nine-week endurance exercise significantly increased the circulating IL-6 concentration compared with sedentary group (239.6 ± 37.2 pg/ml vs. 151.8 ± 31.5 pg/ml, P < 0.01). Exercise rats showed significant increases in JAK2, STAT3, Akt, ERKs, and SOCS3 phosphorylations compared with sedentary rats (P < 0.01). CONCLUSION: The data suggest that endurance exercise is a leptin signaling mimetic in hypothalamus of Wistar rats.

[Treatment of calcaneal avulsion fractures with twinfix suture anchors fixation].

OBJECTIVE: For the calcaneal avulsion fracture, the current method is more commonly used screws or Kirschner wire to fix fracture fragment. This article intended to explore the feasibility and clinical efficacy for the treatment of avulsion fractures with TwinFix suture anchors. METHODS: From July 2007 to November 2010, 21 patients were reviewed, including 15 males and 6 females, ranging in age from 49 to 65 years,with a mean of 58.7 years. Twelve patients had nodules in the right heel and 9 patients had nodules in the left heel. All the patients had closed fractures. The typical preoperative symptoms of the patients included pain in the upper heel and weak in heel lift. Body examination results: palpable sense of bone rubbing in the back of the heel, and swelling in the heel. Surgery treatment with TwinFix suture anchors performed as follows : to fix TwinFix suture anchors into the calcaneal body, then to drill the fracture block, to make the double strand suture through the fracture holes, to knot the suture eachother to fix the block, and to use stitch to fix the remaining suture in the Achilles tendon in order to improve the block fixation. The criteria of the AOFAS Foot and Ankle Surgery by the United States Association of ankle-rear foot functional recovery was used to evaluate the Achilles tendon. RESULTS: Total average score was (95.5 +/- 3.12) points, including pain items of(38.5 +/- 2.18) points,the average score of functional items of (49.5 +/- 3.09) points,and power lines of 10 points in all patients. Twenty-one patients got an excellent result, 16 good and 5 poor. CONCLUSION: The methods of treatment for the calcaneal avulsion fractures with TwinFix suture anchors is a simple operation, and have excellent clinical effect, which is worthy of promotion.

Inhibiting gastric cancer-associated angiogenesis by CIAPIN1 siRNA.

Angiogenesis plays an essential role in tumor growth and metastasis and is a promising target for cancer therapy. We characterized the effects of selective CIAPIN1 inhibition on the angiogenesis gastric cancer cell line SGC7901 by stable transfection of CIAPIN1 siRNA. Our study has been shown that CIAPIN1 play the determined role in tumor growth and multidrug resistance. The conditioned media obtained from SGC7901 treated with CIAPIN1 siRNA suppressed in vitro the proliferation, migration and tube formation of human umbilical vein endothelial cells compared with untransfected cells or cells transfected with control vector alone. Furthermore, the stable transfection of CIAPIN1 siRNA inhibited in vivo tumorigenicity and angiogenesis. Our findings support that selective inhibition of CIAPIN1 alone plays an instrumental role on gastric cancer associated angiogenesis.

Clinical effects of acupuncture after surgical operation in patients with prolapse of the lumbar intervertebral disc.

OBJECTIVE: To investigate the clinical effects of acupuncture after surgical operation in patients with prolapse of the lumbar intervertebral disc (PLID). METHODS: Sixty-nine patients in this series, who had undergone the removal of nucleus pulposus and the intervertebral fusion as well, were randomly divided into a treatment group of 35 cases and a control group of 34 cases. The former was treated by acupuncture and conventional rehabilitation therapy, and the latter only by the rehabilitation therapy. The therapeutic effects were evaluated according to the scoring system stipulated by Japanese Orthopedics Association (JOA). RESULTS: In the treatment group, the average functional recovery rates in 3-month, 6-month and one-year periods were respectively 49.93%, 90.31% and 95.08%; while the rates were repesctively 26.24%, 63.42% and 71.36% in the control group, showing statistically significant difference between the two groups (P<0.05). CONCLUSIONS: Acupuncture can confirmatively promote the functional recovery for'patients with prolapse of the lumbar intervertebral disc after surgical removal of nucleus pulposus and with intervertebral fusion.

[The study for the immuno-protection of E. coli O157:H7 outer membrane protein].

OBJECTIVE: To evaluate the protective efficacy against a lethal dose of E. coli O157:H7 after intranasal, oral and subcutaneous immunization with the outer membrane protein (OMP) extracted from the whole cell of E. coli O157:H7. METHODS: Female BALB/C mice were immunized three times (on days 0, 7 and 14) with OMP and CT or Complete Frund adjuvant. On the 21st day after the last immunization, serum, fecal extracts and vaginal washes were collected for the detection of antigen-specific antibody responses by ELISA before the oral challenge with E. coli O157:H7 933. And the antigens that induced the specific antibody responses were analysed by Western-blotting. Then, the mice were orally challenged with 0.3 ml 4.25 x 10(10)/ml live E. coli O157:H7, and the mortality was recorded. On the 7th day after the challenge, the mice were sacrificed and the heart, liver, lung, kidney, small intestine and colon were collected. Then the histology lesions were observed by light microscopy. RESULTS: In ELISA, both intranasal and oral immunization, with CT as a mucosal adjuvant, induced strong anti-OMP IgA responses in serum, fecal extract and vaginal washes, and anti-OMP IgG responses in serum. However, the intranasal immunization was much more effective to induce specific IgA and IgG responses than the oral immunization. In contrast to mucosal immunization, subcutaneous immunization only induced high levels of specific IgG antibodies in serum, and did not effectively promote IgA immune response. The results of the protective efficacy after challenge showed that both intranasal and oral immunizations with OMP provided significant protection (86.7% to 40%, P < 0.01 and 73.3% to 40%, P < 0.05) against a lethal dose of E. coli O157:H7 challenge, and intranasal immunization possessed a better protective ability (86.7% to 73.3%, P < 0.05). In contrast, the mice immunized subcutaneously were not protected. They were died more early after oral challenge. Furthermore, the histopathological features of the kidney, colon and other organs also appeared to be well correlated with immunization route. In comparison with the mice immunized subcutaneously and those in control group, the severity of the histopathological lesions in the tissues of the mice immunized intranasally was minimal. CONCLUSION: These results suggested that the intranasal immunization should be the best choice of vaccine development against E. coli O157:H7.