Effect of daily milk supplementation on serum and umbilical cord blood folic acid concentrations in pregnant Han and Mongolian women and birth characteristics in China.

Many studies have demonstrated the efficacy of folic acid (FA) supplementation in prevention of neural tube defects (NTDs), although the extent of NTDs varies among individuals of different races and ethnic origin. China is a multi-ethnic country with no standard practice for FA-fortified food. Milk is consumed by women, but little is known about the effects of milk on folate concentration in maternal blood and neonatal umbilical cord blood in Han and Mongolian women after stopping taking the supplement for a month and five month, respectively. The objective of this study was to determine whether only daily consumption of liquid milk can increase the blood folate concentration in pregnant women and whether there are differences in blood folate concentrations between Han and Mongolian women after cessation of FA supplementation. Of the 4052 women enrolled in the parallel group design study. Three thousand five hundred and twenty-six women had confirmed pregnancies and were randomized to receive liquid milk or not until delivery. Women who consumed the liquid milk had significantly increased serum folate concentrations at 16 and 32 weeks of gestation as well as cord blood at birth compared to control groups in both ethnic groups. Infants born to women drinking milk also had better the term birth weight and height, which may be related to the increased concentration of folate. In conclusion, daily consumption of milk can increase the serum folate concentration in pregnant Han and Mongolian women in China (differences in the efficacy of FA and milk supplementation) and may enhance birth outcomes.

Treatment of complicated tibial plateau fractures with dual plating via a 2-incision technique.

The operative treatment of complicated bicondylar fractures of the tibial plateau remains a challenge to most surgeons. This retrospective study was designed to evaluate the clinical and radiological outcomes of dual plating via a 2-incision technique for the repair of complicated bicondylar tibial plateau fractures. A series of consecutive patients with bicondylar tibial plateau fractures treated by open reduction and internal fixation with a double buttress plate or a combination of locking plate and buttress plate via a 2-incision technique between March 2004 and March 2008 were retrospectively analyzed. Radiological and clinical results and complications of the 2 different fixation methods were compared. Seventy-nine patients matching the criteria of this study were followed up for at least 24 months. All of the fractures healed, with 3 cases of deep infection, 7 cases of secondary loss of reduction, 3 cases of secondary loss of alignment, and 10 cases of knee instability. At 24-month follow-up, mean Hospital for Special Surgery scores were 77.8±9.4 and 79.0±7.9 in the double buttress plate group and combination group, respectively. No significant differences in clinical or radiographic outcomes were found between the 2 groups, except that the combination group needed less bone graft. Dual plating with 2 incisions provided good exposition for the reduction and fixation of complicated bicondylar tibial plateau fractures. Using a combination of locking plate and buttress plate reduced the amount of bone graft compared with the double buttress plate technique.

BLCAP induces apoptosis in human Ewing's sarcoma cells.

Bladder cancer-associated protein (BLCAP) is a novel candidate tumor suppressor gene identified from human bladder carcinoma and highly associated with the invasion of bladder cancer. We previously reported that it also plays a key role in the tumorigenesis and metastasis of human osteosarcoma. In the present study, we constructed a recombinant encoding BLCAP cDNA. Overexpression of BLCAP resulted in growth inhibition and induced apoptosis of human TC-135 Ewing's sarcoma cells in vitro. We further investigated the caspase-3/7 activity and expressions of the fusion transcription factor Ewing's sarcoma protein-friend leukemia virus integration 1 (EWS-FLI1) and the apoptosis regulator B-cell lymphoma 2 (BCL-2). Cell apoptosis was accompanied by the down-regulated expression of EWS-FLI1 and BCL-2. Our present results suggest that BLCAP may play a role not only in regulating cell proliferation but also in coordinating apoptosis through the down-regulation of BCL-2 and EWS-FLI1 in human Ewing's sarcoma cells.

Application of locking plate in long-bone atrophic nonunion following external fixation.

The treatment of atrophic fracture nonunion continues to represent a therapeutic challenge. Large segmental osteopenia is often seen in patients who received uniplanar or hybrid external fixators as the definitive method of fixation for high-energy fractures, and this adds more difficulties to the treatment of fracture nonunion. This retrospective study was designed to assess the outcome of locking compression plating with autologous bone grafting in patients with long-bone atrophic nonunion following external fixation.From January 2004 to December 2009, a series of consecutive patients with atrophic nonunion of the long bone following external fixation were treated with this method in our institution. The clinical outcomes and complications of these patients were retrospectively analyzed. Twenty-seven patients with 28 fracture nonunions were involved in this study. Mean follow-up was 14.2±3.4 months. Bony union was achieved in all 27 patients within a mean 18.6±4.8 weeks after revision surgery. Two patients developed superficial wound infections. No deep infections were found, and no implant failure was seen. Three patients reported minor pain in the donor site of the bone graft, and no other donor site complications were found.Revision osteosynthesis of long-bone atrophic nonunion following external fixation by locking compression plating with autologous iliac crest bone grafting represents a safe and efficacious modality for the treatment of these challenging conditions.

Death of axotomized retinal ganglion cells delayed after intraoptic nerve transplantation of olfactory ensheathing cells in adult rats.

Intraorbital transection of the optic nerve (ON) always induces ultimate apoptosis of retinal ganglion cells (RGCs) and consequently irreversible defects of vision function. It was demonstrated that transplanted olfactory ensheathing cells (OECs) in partially injured spinal cord have a distant in vivo neuroprotective effect on descending cortical and brain stem neurons. However, this study gave no answers to the question whether OECs can protect the central sensitive neurons with a closer axonal injury because different neurons respond variously to similar axonal injury and the distance between the neuronal soma and axonal injury site has a definite effect on the severity of neuronal response and apoptosis. In the present study, we investigated the effect of transplanted OECs on RGCs after intraorbital ON transection in adult rats. Green fluorescent protein (GFP)-OECs were injected into the ocular stumps of transected ON and a significantly higher number of surviving RGCs was found together with a consistent marked increase in the mRNA and protein levels of BDNF in the ON stump and retina in the OEC-treated group at 7 days, but not 2 and 14 days, time point when compared to the control group. Our findings suggest that OEC transplantation induces the expression of BDNF in the ocular ON stump and retina and delays the death of axotomized RGCs at a certain survival period.

[ShRNA of Cyclin D1 decreased the proliferation of human osteosarcoma cell line SOSP-9607.].

AIM: To investigate the effect of Cyclin D1 shRNA on the apoptosis and proliferation of human osteosarcoma cell line SOSP-9607. METHODS: Human Cyclin D1 shRNA vector was stably transfected into SOSP-9607 osteosarcoma cells. The mRNA and protein cxpression levels of Cyclin D1 were detected by semiquantitative RT-PCR and Western blot respectively. The cell cycle and pretiferation of osteosarcoma cells were examined by FCM analysis and CCK-8 method, respectively. RESULTS: After stable transfection of Cyclin D1 shRNA, the expression of Cyclin D1 were inhibited at mRNA and protein levels. The proliferation of SOSP-9607 osteosarcoma cells was inhibited. The difference was significant compared with control groups (P<0.05). At the same time, Cyclin D1 shRNA transfection increased G0/1 phage content and decreased S phage content. CONCLUSION: Cyclin D1 shRNA could down-regulate the expression of Cyclin D1, effectively inhibit the proliferation of osteosarcoma cells, and have significant effect on the cell cycle.

Silencing of calpain expression reduces the metastatic potential of human osteosarcoma cells.

Osteosarcoma, the most common primary bone tumor in young adults, is characterized by local invasion and distant metastasis. But detailed mechanisms of tumorigenicity and metastasis of osteosarcoma are not well known. We report the involvement of calpains, a family of calcium-activated, cysteine proteases, in the invasive and metastatic processes of human osteosarcoma cells. By using siRNA treatment, the expression of mu- and m-calpains were downregulated in human Saos-2 osteosarcoma cells. Both the adhesive and invasive potentials were significantly attenuated in calpain siRNA-transfected human Saos-2 osteosarcoma cells. MMPs are the main factors involved in malignant tumor invasion and metastasis. siRNA of calpains also significantly inhibited the secretion of MMP-2 in Saos-2 cells. These results suggest that mu- and m-calpains are important in the invasion and metastasis of human osteosarcoma cells, and calpains might be targeted to reduce tumor progression.

Mu-calpain is involved in the regulation of TNF-alpha-induced matrix metalloproteinase-3 release in a rheumatoid synovial cell line.

Calpain is secreted by intra-articular synovial cells and degrades the main components of cartilage matrix proteins, proteoglycan, and collagen, causing cartilage destruction. Matrix metalloproteinase-3 (MMP-3) has also been detected in synovial fluid and serum, and is involved in the development and progression of rheumatoid arthritis by degradation of the extracellular matrix and cartilage destruction. To investigate the relationship between calpain and MMP-3 in rheumatic inflammation, we utilized the rheumatic synovial cell line, MH7A. Tumor necrosis factor (TNF-alpha) stimulation-induced increased expression of mu-calpain, m-calpain, and MMP-3 in these cells, as well as the release of calpain and MMP-3 into the culture medium. The calpain inhibitors, ALLN (calpain inhibitor I) and calpeptin, did not affect the intracellular expression of MMP-3, but reduced the secretion of MMP-3 in a concentration-dependent manner. Down-regulation of mu- but not m-calpain by small interfering RNAs abolished TNF-alpha-induced MMP-3 release from the synovial cells. These findings suggest that calpain, particularly mu-calpain, regulates MMP-3 release by rheumatic synovial cells, in addition to exerting its own degradative action on cartilage.

Inhibition of platelet-derived growth factor-induced cell growth signaling by a short interfering RNA for EWS-Fli1 via down-regulation of phospholipase D2 in Ewing sarcoma cells.

EWS-Fli1, a fusion gene resulting from a chromosomal translocation t(11;22, q24;q12) and found in Ewing sarcoma and primitive neuroectodermal tumors, encodes a transcriptional activator and promotes cellular transformation. However, the precise biological functions of its products remain unknown. To investigate the role of EWS-Fli1 in cell growth signaling, we transfected Ewing sarcoma TC-135 cells with short interfering RNAs for EWS-Fli1. EWS-Fli1 knockdown reduced cell growth and platelet-derived growth factor (PDGF)-BB-induced activation of the growth signaling enzymes. Interestingly, phospholipase D2 (but not the PDGF-BB receptor) showed marked down-regulation in the EWS-Fli1-knocked down TC-135 cells compared with the control cells. In Ewing sarcoma TC-135 cells, the PDGF-BB-induced phosphorylation of growth signaling involving extracellular signal-regulated kinase, Akt, p70S6K, and the expression of cyclin D3 were markedly inhibited by transfection with short interfering RNA phospholipase (PL)-D2. The PDGF-BB-induced activation of growth signaling was also suppressed by 1-butanol, which prevents the production of phosphatidic acid by phospholipase D (but not by t-butyl alcohol), thereby implicating PLD2 in PDGF-BB-mediated signaling in TC-135 cells. These results suggest that EWS-Fli1 may play a role in the regulation of tumor proliferation-signaling enzymes via PLD2 expression in Ewing sarcoma cells.

[Relationship between expression of BLCAP protein and malignancy of osteosarcoma].

AIM: To study the relationship between expression of bladder cancer-associated protein (BLCAP) and malignancy of osteosarcoma. METHODS: SABC method was applied to study the expression of BLCAP protein in osteosarcoma, according to clinical results, to analyze the prognosis value of BLCAP protein in osteosarcoma. RESULTS: The positive rate of BLCAP protein in primary osteosarcoma was 65.6%,much higher than that in recurrent osteosarcoma, which was 25.0%. CONCLUSION: It may be helpful for evaluating the prognosis of osteosarcoma to study the relationship between expression of BLCAP protein and malignancy of osteosarcoma.

[Construction and characterization of osteosarcoma 9901 cell cDNA library].

AIM: To construct human osteosarcoma 9901 cell cDNA expression library for screening osteosarcoma-specific antigens. METHODS: Total RNA was extracted from human osteosarcoma cell line 9901 and mRNA was purified. cDNA was synthesized by reverse transcription, and cDNA fragments larger than 400 bp were ligated with dephosphorylated arms of lambdagt11. The recombinants were packaged in-vitro, and a small portion of packaged phage was used to infect E.coli Y1090 for titration. The size of cDNA inserts and the diversity of library were detected by PCR. RESULTS: The osteosarcoma 9901 cell line cDNA library consisting of 1.5x10(6) recombinant bacteriophages was constructed. The average length of exogenous inserts in the recombinants was about 1.4 kb. CONCLUSION: The cDNA library reported herein is suitable for screening osteosarcoma-specific antigens.