Objective To investigate the effect of different concentrations of raloxifene (RAL) on the proliferation and apoptosis of human aortic valve interstitial cells (AVICs) in vitro. Methods AVICs were isolated from human aortic valve by collagenase type Ⅱ, and cultured in different concentrations (0 nmol/L, 0.1 nmol/L, 1 nmol/L,10 nmol/L, 100 nmol/L and 1 000 nmol/L) of RAL. AVICs cultured in 0 nmol/L RAL were treated as the control group and those in other concentrations of RAL as the experiment groups. The proliferation and apoptosis of AVICs were evaluated by Cell Proliferation Assay (MTS assay) on day 0, 3, 5, 7 and 9. Flow cytometry was used to detect the cell cycle and apoptosis of AVICs on day 7. Results MTS results showed that the optical density value at 490 nm was much less in 10 nmol/L RAL and 100 nmol/L RAL groups (P<0.05) on day 5, 7 and 9 than that in the control group. Flow cytometry results demonstrated that S-phase rate (P<0.05) and cell apoptosis rate (P<0.05) on day 7 were lower in the 10 nmol/L and 100 nmol/L RAL groups compared with the control group. Conclusion RAL with suitable concentration can inhibit proliferation and apoptosis of AVICs, which will lay an important foundation for further research of the role of RAL on heart valve diseases.
Objective To investigate the way to culture human parathyroid cells and to investigate its secretory function. Methods After digested by collagenase, parathyroid cells were isolated to get the original generation cells, then the cells were cultured and passaged, and morphological changes of original generation cells and passage cells were observed on every day. The parathyroid hormone(PTH) level secreted by the original generation cells and passage cells were measured on the 1st, 5th, 10th, 15th, and 20th day(original generation cells only) respectively. Results The cellular morphology was complete after digestion. On the 2nd day, most of the parathyroid cells had adhered and spreaded, on the 3rd day, all cells had spread. There was no very obvious changes on these cells after cultured for 4-15 days. From 16 to 20 days, some parathyroid cells went senescence. On the 1st day, all of the passage cells, which were fusiform and little bigger than those of the original generation cells, had adhered and spreaded. From 2 to 15 days, there was no very obvious changes. The concentration of PTH in original generation cells begin to decreased significantly on the 10th day (P < 0.01). The concentration of PTH in passage cells were all lower than those of original generation cells at the same corresponding time, but there were no significant difference on the PTH level on 5th day and 1st day, 10th day and 5th day, 15th day and 10th day in passage cells (P > 0.05). Conclusion Parathyroid cells which were cultured within 10 days possess well morphologic structure and have the strongest secretory function. Although the passage cells still possess secretory function, it is greatly inferior to original generation cells. At last, we consider that original generation cells cultured within 10 days can be regarded as the source of allogeneic cell transplantation.
Objective To investigate the reliability of culture method of adult human parathyroid cells. Methods Adult human parathyroid tissue was digested by collagenase, then the original generation of cells were cultured and passaged, and their morphological changes were observed and recorded every other day. Part of the passaged cells were observed through electron microscope and its supernatant parathyroid hormone (PTH) was assayed. Meanwhile, the other part of cells were tested the parathyroid markers, including PTH, calcium-sensing receptor (CaSR) and glial cells missing-2 (GCM-2) by PCR. Results Abundant cytoplasm, mitochondria, endoplasmic reticulum and Golgi apparatus from the seventh day's passaged cells were observed by the electron microscopy, as well as, some secretory granules existing in both cytoplasm and intercellular lacuna. Also, the PTH from supernate was detected, and parathyroid specific markers, such as CaSR, PTH, and GCM2 were positive. Conclusions These trials demonstrated the adult human parathyroid cells could be harvested by collagenase digestion and the cultured. Furthermore, the cells remained good shape and kept functioning, making it a potential source for allogeneic cell transplantation to the treatment of permanent hypoparathyroidism.
ObjectiveTo systematically review the efficacy of at the fracture level (AFL) versus cross the fracture level (CFL) short-segment pedicle screw fixation for thoracolumbar fractures. MethodsWe electronically search PubMed, The Cochrane Library (Issue 8, 2015), EMbase, CBM, CNKI, VIP and WanFang data to collect randomized controlled trials (RCTs) of AFL versus CFL short segment pedicle screw fixation for thoracolumbar fractures from inception to Aug. 2015. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Meta-analysis was performed using RevMan 5.3 software. ResultsA total of 11 RCTs involving 730 patients were included. The results of meta-analysis indicated that: compared with the CFL group, the AFL group had more blood loss (MD=9.8, 95%CI 7.40 to 12.20), less implant failure rate (RR=0.19, 95%CI 0.07 to 0.48), lower long term postoperative VAS score of thoracolumbar pain (MD=-1.20, 95%CI -1.85 to -0.56), higher correction in short term postoperative kyphotic Cobb angle (MD=3.56, 95%CI 2.25 to 4.87), smaller value in long term postoperative kyphotic Cobb angle and its loss of correction (MD=-3.95, 95%CI -7.78 to -0.12; MD=-4.65, 95%CI -6.91 to -2.40), smaller degree of anterior vertebral height compression in short and long term postoperative (MD=-3.51, 95%CI -5.23 to -1.80; MD=-8.28, 95%CI -12.22 to -4.33), better result in long term postoperative anterior vertebral height and its loss of correction (MD=8.00, 95%CI 3.85 to 12.15; MD=-6.06, 95%CI -7.68 to -4.44). There were no significant differences between two groups regarding operation time, infectious complications and short term postoperative kyphotic Cobb angle (MD=0.11, 95%CI -5.36 to 5.57; RR=0.55, 95%CI 0.11 to 2.85; MD=-0.66, 95%CI -2.19 to 0.87). ConclusionCurrent evidence shows that AFL short-segment pedicle screw fixation for thoracolumbar fractures is superior to CFL fixation. Due to limited quality and quantity of the included studies, more high quality studies are needed to verify the above conclusion.
ObjectiveTo systematically review the effects of autograft versus allograft tendon for posterior cruciate ligament single-bundle reconstruction. MethodsDatabases including PubMed, The Cochrane Library (Issue 3, 2015), EMbase, CBM, CNKI, VIP and WanFang Data were searched from inception to August 2015, to collect randomized controlled trials, clinical controlled trials and cohort studies of autograft tendon versus allograft tendon for posterior cruciate ligament single-bundle reconstruction. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Meta-analysis was performed using RevMan 5.3 software. ResultsA total of 7 cohort studies involving 376 patients who had undergone the arthroscopic transtibial single-bundle PCL reconstruction were included. The results of meta-analysis indicated that no significant differences were found between the autograft group and the allograft group in Lysholm score (MD=-0.54, 95%CI -2.36 to 1.27, P=0.56), Tegner score (MD=-0.04, 95%CI -0.88 to 0.80, P=0.93), IKDC objective score (OR=1.31, 95%CI 0.68 to 2.53, P=0.41) and posterior translation side-to-side difference (SMD=-0.15, 95%CI -0.37 to 0.07, P=0.18). However, patients in the allograft group had a longer duration of fever when compared with the autograft group patients (MD=-3.55, 95%CI-5.61 to -1.49, P=0.0007). ConclusionCurrent evidence shows that autograft tendon and allograft tendon tibial have similar effects in PCL single-bundle reconstruction, though there is a longer duration of fever in patients with allograft. Due to limited quality and quantity of the included studies, more high quality studies are needed to verify the above conclusion.