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Objective:To investigate the effect of mechano-growth factor（MGF）on osteoclast activity and its mechanism.Methods:The RAW264.7 precursor osteoclast cell line was cultured with 25 ng/ml macrophage-colony stimulating factor（M-CSF）and 30 ng/ml receptor activator of NF-κB ligand（RANKL），and identified by tartrate resistant acid phosphatase（TRAP）staining after 7 days of culture. Western blot anslysis was used to determine the effect of 45 ng/ml MGF on the phosphoinositide-3-kinase/protein kinase B（PI3K/AKT）signaling pathway in separated osteoclasts，including levels of AKT，phosphorylation（p）-AKT，lactation mammalian target of rapamycin（mTOR），p-mTOR and TRAP at 0，4，8 and 12 hours. Real-time fluorescence quantitative PCR was used to expressions of TRAP in osteoclasts at 0，4，8 and 12 hours. The PI3K/Akt phosphorylation inhibitor LY294002（20 μmol/L）combined with MGF（45 ng/ml）was used to act on osteoclasts，and expression levels of Akt，p-Akt，mTOR，p-mTOR and TRAP were detected by Western blot at 0，4，8 and 12 hours.Results:After culturing RAW264.7 cells with M-CSF and RANKL for 7 days，a large number of osteoclasts with positive TRAP staining can be obtained. Western blot analysis showed expression levels of Akt and mTOR did not change significantly over time（ P>0.05），expression levels of p-Akt and p-mTOR increased continuously from（2.18±0.34）pg/ml and（0.83±0.10）pg/ml at 0 hour to（3.86±0.36）pg/ml and（1.56±0.19）pg/ml at 12 hours（ P<0.05），and expression level of TRAP decreased significantly over time，from（5.66±0.47）pg/ml at 0 hour to（3.76±0.38）pg/ml at 12 hours（ P<0.05）. Real-time fluorescence quantitative PCR analysis of expression of TRAP in osteoclasts showed that MGF inhibited the expression of TRAP in osteoclasts，which decreased from 1.02±0.06 at 0 hour to 0.53±0.11 at 12 hours（ P<0.05）. After acting LY294002 combined with MGF on osteoclasts，Western blot analysis showed expression levels of Akt and mTOR did not change significantly over time（ P>0.05），expression levels of p-AKT and p-mTOR decreased significantly from（3.28±0.18）pg/ml and（3.29±0.22）pg/ml at 0 hour to（2.06±0.34）pg/ml and（2.04±0.20）pg/ml at 12 hours（ P<0.05），and expression level of TRAP had no significant difference over time（ P>0.05）. Conclusions:MGF inhibits osteoclast activity by inhibiting the expression of TRAP in osteoclasts through PI3K/Akt signaling pathway. LY294002 inhibits the expression of PI3K/Akt signaling pathway in osteoclasts，further verifying the mechanism of MGF inhibiting osteoclast activity，and this finding puts forward new ideas for clinical prevention and treatment of osteoporosis.
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Objective To evaluate combined robotic and endoscopic surgery in the third space for gastric submucosal tumors (SMTs).Methods Combined surgery in 4 patients were compared with 19 patients who underwent laparoscopic wedge resection between Aug 2017 and Feb 2018.Results R0 resection was achieved in all combined surgery patients.The operation time was longer (112 ±29)min vs.(93 ±11) min (t =2.338,P＜0.05),but less blood loss,(11 ±6)ml vs.(59 ±6)ml (t =15.102,P＜ 0.01).In combined surgery group,there was a larger tumor over resected tissue percentage,65.4％ ± 28.2％ vs.22.8％ ± 19.6％ (t =3.680,P ＜ 0.05).Combined surgery group patients were with earlier first breaking wind (3 ± 1.4) d vs.(4.1 ± 0.9) d (t =2.026,P ＞ 0.05),and shorter hospital stay (6.5 ± 1.3) d vs.(8.5 ±0.6) d (t =4.902,P＜0.01),but a little higher hospitalization costs(57 651 ± 10 097) rmb vs.(42 620 ± 5 086) rmb (t =4.508,P ＜ 0.01).There were no major postoperative complications occurred in neither groups,nor tumor recurrences after 3-month follow-up.Conclusion Combined robotic and endoscopic surgery in the third space for submucosal gastric tumors is a novel operation with short operation time,less blood loss and good result.
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Objective@#To evaluate combined robotic and endoscopic surgery in the third space for gastric submucosal tumors(SMTs).@*Methods@#Combined surgery in 4 patients were compared with 19 patients who underwent laparoscopic wedge resection between Aug 2017 and Feb 2018.@*Results@#R0 resection was achieved in all combined surgery patients. The operation time was longer (112±29 )min vs. (93±11 ) min (t=2.338, P<0.05), but less blood loss, (11±6)ml vs.(59±6)ml (t=15.102, P<0.01). In combined surgery group, there was a larger tumor over resected tissue percentage, 65.4%±28.2% vs.22.8%±19.6% ( t=3.680, P<0.05). Combined surgery group patients were with earlier first breaking wind (3±1.4) d vs.(4.1±0.9) d (t=2.026, P>0.05), and shorter hospital stay (6.5±1.3) d vs.( 8.5±0.6) d (t=4.902, P<0.01), but a little higher hospitalization costs(57 651±10 097)rmb vs.(42 620±5 086)rmb (t=4.508, P<0.01). There were no major postoperative complications occurred in neither groups, nor tumor recurrences after 3-month follow-up.@*Conclusion@#Combined robotic and endoscopic surgery in the third space for submucosal gastric tumors is a novel operation with short operation time, less blood loss and good result.
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Objective To explore the effects of bone marrow mesenchymal stem cells (BMSCs) in different concentrations on the balance of regulatory T cell / T-helper cell 17 (Treg / Th17). Methods BMSCs were isolated from SPF grade male Wistar rats with age of 3 weeks old and weight of 50 g. BMSCs were cultured and identified when they were expanded to the 4th generation. CD4+ T lymphocytes were isolated from SPF grade male Wistar rat with age of 6 weeks old and weight of 200 g and assayed for cell purity by flow cytometry. BMSCs were divided into 0.5-fold concentration group, basal concentration group, 2-fold concentration group and 4-fold concentration group by their concentrations of 1×105/well, 2×105/well, 4×105/well and 8×105/well, which were cultured with CD4+ T lymphocytes for 72 hours, respectively. Then the proportion of Treg cells and Th17 cells in each group was detected by flow cytometry, and cytokines were detected by cytometric bead array. Results The purities of BMSCs and CD4+ T lymphocytes were both higher than 95%. In the co-culture of BMSCs and CD4+ T lymphocytes, the proportions of Treg cells were statistically different among different concentration groups of BMSCs (F = 10.071, P = 0.001), in which BMSCs in 2-fold concentration group had the strongest ability to promote the Treg cells proliferation. The proportion of Treg cells in 2-fold concentration group was significantly higher than that in 0.5-fold concentration group, basal concentration group and 4-fold concentration group [(9.24±2.68)% vs. (3.87±0.38)%, (5.16±1.69)%, (3.86±0.36)%, all P < 0.01]. The level of interleukin-10 (IL-10) was lowest in 0.5-fold concentration group, and it was significantly lower than that in basal concentration group, 2-fold concentration group and 4-fold concentration group (ng/L: 39.80±14.48 vs. 148.43±64.49, 156.40±59.27, 126.92±42.95, all P < 0.05). Transforming growth factor-β (TGF-β) was the highest in basal concentration group, and it was significantly higher than that in 0.5-fold concentration group, 2-fold concentration group and 4-fold concentration group [ng/L: 3.17 (1.88, 5.74) vs. 0.71 (0.32, 1.38), 1.22 (0.47, 2.97), 0.52 (0.37, 1.23), all P < 0.05]. The proportions of Th17 cells were statistically different among the different concentration groups (F = 21.069, P = 0.000), with the highest proportion in basal concentration group which was significantly higher than that in 0.5-fold concentration group or 4-fold concentration group [(0.89±0.08)% vs. (0.64±0.15)%, (0.37±0.10)%, both P < 0.01], but no significant difference was found as compared with 2-fold concentration group [(0.83±0.06)%, P > 0.05]. However, the expressions of IL-17 and IL-6 were not different among the different concentration groups respectively (IL-17: χ2 = 0.550, P = 0.760;IL-6: χ2 = 0.010, P = 0.995). Conclusions BMSCs in moderate concentrations [(2-4)×105/well] could promote proliferation both in Treg cells and Th17 cells, but no change could be found in higher concentrations of BMSCs (8×105/well). However, the changes in related cytokines were not synchronized with Treg / Th17 cells.
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OBJECTIVE@#To explore the effects of bone marrow mesenchymal stem cells (BMSCs) in different concentrations on the balance of regulatory T cell/T-helper cell 17 (Treg/Th17).@*METHODS@#BMSCs were isolated from SPF grade male Wistar rats with age of 3 weeks old and weight of 50 g. BMSCs were cultured and identified when they were expanded to the 4th generation. CD4+ T lymphocytes were isolated from SPF grade male Wistar rat with age of 6 weeks old and weight of 200 g and assayed for cell purity by flow cytometry. BMSCs were divided into 0.5-fold concentration group, basal concentration group, 2-fold concentration group and 4-fold concentration group by their concentrations of 1×105/well, 2×105/well, 4×105/well and 8×105/well, which were cultured with CD4+ T lymphocytes for 72 hours, respectively. Then the proportion of Treg cells and Th17 cells in each group was detected by flow cytometry, and cytokines were detected by cytometric bead array.@*RESULTS@#The purities of BMSCs and CD4+ T lymphocytes were both higher than 95%. In the co-culture of BMSCs and CD4+ T lymphocytes, the proportions of Treg cells were statistically different among different concentration groups of BMSCs (F = 10.071, P = 0.001), in which BMSCs in 2-fold concentration group had the strongest ability to promote the Treg cells proliferation. The proportion of Treg cells in 2-fold concentration group was significantly higher than that in 0.5-fold concentration group, basal concentration group and 4-fold concentration group [(9.24±2.68)% vs. (3.87±0.38)%, (5.16±1.69)%, (3.86±0.36)%, all P < 0.01]. The level of interleukin-10 (IL-10) was lowest in 0.5-fold concentration group, and it was significantly lower than that in basal concentration group, 2-fold concentration group and 4-fold concentration group (ng/L: 39.80±14.48 vs. 148.43±64.49, 156.40±59.27, 126.92±42.95, all P < 0.05). Transforming growth factor-β (TGF-β) was the highest in basal concentration group, and it was significantly higher than that in 0.5-fold concentration group, 2-fold concentration group and 4-fold concentration group [ng/L: 3.17 (1.88, 5.74) vs. 0.71 (0.32, 1.38), 1.22 (0.47, 2.97), 0.52 (0.37, 1.23), all P < 0.05]. The proportions of Th17 cells were statistically different among the different concentration groups (F = 21.069, P = 0.000), with the highest proportion in basal concentration group which was significantly higher than that in 0.5-fold concentration group or 4-fold concentration group [(0.89±0.08)% vs. (0.64±0.15)%, (0.37±0.10)%, both P < 0.01], but no significant difference was found as compared with 2-fold concentration group [(0.83±0.06)%, P > 0.05]. However, the expressions of IL-17 and IL-6 were not different among the different concentration groups respectively (IL-17: χ2 = 0.550, P = 0.760; IL-6: χ2 = 0.010, P = 0.995).@*CONCLUSIONS@#BMSCs in moderate concentrations [(2-4)×105/well] could promote proliferation both in Treg cells and Th17 cells, but no change could be found in higher concentrations of BMSCs (8×105/well). However, the changes in related cytokines were not synchronized with Treg/Th17 cells.
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Animales , Masculino , Ratas , Citocinas/metabolismo , Células Madre Mesenquimatosas/metabolismo , Ratas Wistar , Linfocitos T Reguladores/metabolismo , Células Th17/metabolismoRESUMEN
Objective To evaluate application of Steinmenn pins to assist reduction in the treatment of unstable intertrochanteric fractures with proximal femoral nail antirotation(PFNA).Methods From February 2010 to June 2013,38 unstable intertrochanteric fractures were treated by us.There were 23 men and 15 women,aged from 32 to 69 years.By Evans-Jensen classification,18 cases were type Ⅲ,13 type Ⅳ and 7 type Ⅴ.They were divided into 2 groups (n =19).Group A received reduction on a traction bed assisted by Steinmenn pins plus PFNA fixation while group B received reduction only on a traction bed plus PFNA fixation.The 2 groups were compared in terms of fracture reduction,operation time,intro-operative blood loss,fracture healing time,and Harris scores one year postoperation.Results According to the evaluation system modified by Baumgaetner et al.,the postoperative quality of fracture reduction was fine in 15 cases and fair in 4 in group A while it was fine in 9 cases,fair in 8 and poor in 2 in group B,showing a significant difference between the 2 groups (P ＜ 0.05).The operation time in group A (50.7 ± 11.9 min) was significantly shorter than in group B (63.4 ± 15.1 min),and the hip joint Harris score (89.4 ±4.4) one year after operation for group A was significantly higher than that for group B (79.6 ±6.4) (P ＜ 0.05).There were no significant differences between the 2 groups regarding intraoperative blood loss,fracture heeling time and follow-up time (P ＞ 0.05).No cases of refracture,delayed union,nonunion,or avascular necrosis of the femoral head were reported.Conclusion In the treatment of unstable intertrochanteric fractures,compared with reduction only on a traction bed plus PFNA fixation,application of Steinmenn pins to assist reduction on a traction bed plus PFNA fixation may lead to better curative efficacy due to its limited invasion,simplicity and beneficial assistance in reduction.
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BACKGROUND:The ankle joint is capable of flexion and extension, including plantar flexion and dorsiflexion, to act as a support and a lever. An ankle injury, often accompanied by fracture and ligament injury, seriously threatens the ankle joint function. Previous diagnosis of ankle injury mainly relied on clinical signs and X-ray examination. However, X-ray examination is not accurate enough for ankle injury diagnosis because it cannot clearly diagnose damage to the surrounding ligaments, tendons and other soft tissues except for obvious fractures. Mutlisequence and multiplanar MRI is currently the optimal noninvasive method for high-resolution determination of soft tissue deformations, but little has been reported on the diagnostic accuracy of ankle ligament and tendon injury. OBJECTIVE:To observe the diagnostic value of MRI for ligament and tendon injury of the ankle in its normal position, and during complete plantar flexion and dorsiflexion. METHODS:It is a single-center, prospective, diagnostic trial that wil be completed at the First Hospital of Hebei Medical University, China. Sixty cases were recruited, including 30 cases of normal ankle joint and 30 cases of ankle ligament and tendon injury. MRI scans of the ankle joint in normal position, complete plantar flexion and complete dorsiflexion were performed in al the cases, and the multi-position MRI results were compared. The primary outcome measure is the sensitivity of MRI to ligament and tendon injury of the ankle during complete plantar flexion. The secondary outcomes include the specificity, positive predictive value, negative predictive value, positive likelihood ratio, and negative likelihood ratio of MRI to ligament and tendon injury of the ankle during the complete plantar position as wel as rate of correct diagnosis;specificity and sensitivity, positive predictive value, negative predictive value, positive likelihood ratio, and negative likelihood ratio in normal position or during the complete dorsiflexion as wel as rate of correct diagnosis;the morphology of the ankle on the multi-position MRI. This study design was registered at ClinicalTrial.gov (03049423) on February 8, 2017. This study protocol has been approved by the Medical Ethics Committee of Qinghai University Affiliated Hospital of China (approval No. 2015076) and wil be performed in accordance with the guidelines of the Declaration of Helsinki, formulated by the World Medical Association. Signed informed consent regarding trial procedure and treatment was obtained from each volunteer. DISCUSSION:This study aims to determine the rate of correct diagnosis of ankle ligament and tendon injury using the MRI, to clarify the diagnostic value of MRI for ankle ligament and tendon injury, and to provide a quantitative MRI diagnostic standard for developing a reasonable surgical treatment.
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Objective To explore the trend of changes of Th1 and Th2 cytokines within 6 months after adult liver transplantation (LT). Methods Twenty-three patients from Tianjin First Center Hospital were chosen as the patient group with an average age of (52.7 ± 7.6), range from 37 to 63 years old, including 21 males and 2 females. Twenty healthy staffs from Tianjin First Center Hospital formed the control group (C) with 15 males and 2 females whose average age was (31.0 ± 6.1) ranged from 22 to 24 years old. The patient group was treated with tacrolimus after LT as main immunosuppressive drug. The peripheral blood at time points before (T0) and 1 month (T1), 3 months (T3), 6 months (T6) after LT at 9:00 AM were collected. The blood sample was also collected form control group but only one time. Levels of IL-2, IFN-γ, IL-10 and TGF-βwere detected by ELISA. Results (1) The concentration of IL-2 showed a continuous up-going trend, which was not such obvious between T1 and T0, and until T3 reached a higher concentration than T0. The concentration at T6 was higher than T0 and T1. There were no significant differences in concentrations of T0 to T3 between patient group and control group, while T6 reached a higher concentration in patient group than that of the control group. (2) The concentration of IFN-γexperienced a shortly down-going trend from T0 to T3, and started rising, reached the peak at 3 months after the operation, then started its down-going trend. There were no significant differences in the concentrations of IFN-γfrom T1 to T6, and T3 reached a higher concentration than T1 while T6 was lower than T3. Only at T3, the concentration of IFN-γwas higher in patient group than that of control group. (3) There were no significant differences in the concentrations of IL-10 at various time points in patient group, and there were no significant differences in the concentrations of IL-10 at different time points between two groups (P>0.05). (4) The concentration of TGF-βshowed a gradual decline after the operation, and reached its bottom at T6, and which was lower than T0 to T3. Compared with the control group, the down-going trend was not such obviously at T0 and T1, and the concentration was down at T3 and T6(P<0.05). Conclusion Our results suggest that there is a tendency of an increasing Th1 cytokine expression at early stage in post-transplantation, while the TGF-βof Th2 cytokine is a decreasing trend. This tendency may associate with the autoimmunity response caused by LT and the immunosuppressive drugs.