1.Effects of microRNA-145 on CD44 expression and hepatocellular carcinoma cells
Linpei WANG ; Xiaoqiu MA ; Weifeng HUANG ; Wei WANG ; Waishi YU ; Jinbin LUO
Chinese Journal of Hepatobiliary Surgery 2019;25(3):179-183
Objective To study the relationship of microRNA-145(miR-145) and CD44 in the hepatocellular carcinoma(HCC).Methods 13 clinical samples of HCC tissues and corresponding normal liver tissues were collected at department of General Surgery,Second Affiliated Hospital of Fujian Medical University from October 2015 to June 2016.The patients with positive expression of CD44 were studied.The expression levels of miR-145 and CD44 in HCC tissues and corresponding normal tissues were detected by real-time quantitative PCR.Western blot was performed to detect the expression of CD44 in HCC cells Hep G2 and SNU423.Biological information methods predicted whether miR-145 and CD44 have a targeted relationship.CD44 expression levels were detected after high expression of miR-145 in HCC cell line with positive expression of CD44.The vector and luciferase reporter genes were constructed to verify the interaction between miR-145 and CD44.The effects of miR-145 on proliferation in HCC cell lines with positive and negative CD44 expression were examined by tetramethylazoazole salt (MTT) assay.Results 8 of the 13 patients showed positive CD44 expression in HCC tissues.Compared with normal liver tissues,the relative expression of miR-145 in HCC tissues was significantly lower (0.998±0.010 vs.0.503±0.046,P<0.05),and the relative expression of CD44 was higher (0.996±0.005 vs.1.878±0.108,P<0.05).Bioinformatics suggested that miR-145 had a targeted relationship with CD44.High expression of miR-145 can significantly reduce the expression level of CD44 mRNA in HCC cell SNU423 (0.941±0.010 vs.0.515±0.021,P<0.05) and down-regulate the expression of CD44 protein.Confirmed by luciferase reporter assay,CD44 is the target gene of miR-145.After transfection with miR-145 mimics,the proliferation of CD44 positive cell SNU423 was significantly inhibited (P<0.05).Conclusions miR-145 can affect the proliferation of CD44 positive HCC cells by regulating the expression of CD44,which may be one of the pathogenesis of HCC.
2.Clinical study on the treatment of diabetic foot ulcer of lateral tibia bone removal microvessel regeneration combined with hyperbaric oxygen
Zhongwei JIA ; Jianping YU ; Han YU ; Yunxing SU ; Jinbin YU ; Zhilan REN ; Jie WEI
Chinese journal of nautical medicine and hyperbaric medicine 2018;25(1):22-26
Objective To investigate clinical efficacy of lateral tibia bone removal micro vessel regeneration combined with hyperbaric oxygen (HBO) in the treatment of diabetic foot ulcer.Methods A retrospective analysis was made on the clinical practice of lateral tibia bone removal micro vessel regeneration combined with HBO in 21 patients (the treatment group)with diabetic foot ulcer who received treatment in the hospital from November 2015 to March 2017.The patients in the treatment group were compared with the 21 patients (the control group) with diabetic foot ulcer who were treated with HBO at the same period.For the patients in the treatment group,a bone window with an area of about 2 cm × 10 cm was performed at medial surface of the affected lateral tibia,and a special external fixation lateral traction frame was placed and debridement was performed on the affected foot at the same time.On the first day after surgery,the patients received HBO therapy with a pressure of 0.20 MPa,once a day,10 days a treatment course,with a total of 3 courses.On the fifty day after surgery,the bone window was slowly removed laterally (1 mm/day).For the patients in the control group,debridement of the affected foot was performed,and on the first day after surgery,HBO therapy was also implemented in the control group with the same treatment scheme as the treatment group.Before surgery and 3 months after surgery,corresponding dada of healing time,ankle brachial index (ABI) and Michigan neuropathy sensation instruments (MNSI) were closely observed in the patients of the 2 groups.Results The patients of the two groups all had medical followed-ups for an average of 9.3 months.In the control group,the wound of one patient with Wagner Ⅴ failed to heal and in the end had amputation,but the wound of the remaining patients all healed,with an average healing time of (39.52 ± 8.43) days.The wound of the treatment group all healed,with an average healing time of (31.74 ± 6.12) days.Three months after surgery,the ABI level of the treatment group increased from pre-surgical level 0.56 ± 0.18 to post-surgical level 0.94 ± O.16,and the MNSI level decreased from pre-surgical level 5.92 ± 1.13 to post-surgical level 4.11 ± 1.25.Three months after surgery,the ABI level of the control group increased from pre-surgical level 0.54 ±0.21 to post-surgical level 0.81 ±0.20,and the MNSI level decreased from pre-surgical level 5.74 ± 1.26 to post-surgical level 4.98 ± 1.34.Statistical significance could be noted in wound healing time when comparisons were made between the two groups.Three months after surgery,statistical significance could also be found in ABI and MNSI levels as compared with those detected before surgery.The treatment group was superior to the control group,also with statistical significance.Conclusions Lateral tibia bone removal micro vessel regeneration could be used to reconstruct vasoganglion below lower leg and promote surrounding vessel and nerve function recovery.Combined with HBO therapy,it could promote wound surface healing,with the advantages of simple surgery,accurate curative effect,high success rate of limb-salvage.For this reason,it was one of the ideal methods for the treatment of diabetic foot ulcer.
3.Clinical study on the treatment of diabetic foot ulcer of lateral tibia bone removal microvessel regeneration combined with hyperbaric oxygen
Zhongwei JIA ; Jianping YU ; Han YU ; Yunxing SU ; Jinbin YU ; Zhilan REN ; Jie WEI
Chinese journal of nautical medicine and hyperbaric medicine 2018;25(1):22-26
Objective To investigate clinical efficacy of lateral tibia bone removal micro vessel regeneration combined with hyperbaric oxygen (HBO) in the treatment of diabetic foot ulcer.Methods A retrospective analysis was made on the clinical practice of lateral tibia bone removal micro vessel regeneration combined with HBO in 21 patients (the treatment group)with diabetic foot ulcer who received treatment in the hospital from November 2015 to March 2017.The patients in the treatment group were compared with the 21 patients (the control group) with diabetic foot ulcer who were treated with HBO at the same period.For the patients in the treatment group,a bone window with an area of about 2 cm × 10 cm was performed at medial surface of the affected lateral tibia,and a special external fixation lateral traction frame was placed and debridement was performed on the affected foot at the same time.On the first day after surgery,the patients received HBO therapy with a pressure of 0.20 MPa,once a day,10 days a treatment course,with a total of 3 courses.On the fifty day after surgery,the bone window was slowly removed laterally (1 mm/day).For the patients in the control group,debridement of the affected foot was performed,and on the first day after surgery,HBO therapy was also implemented in the control group with the same treatment scheme as the treatment group.Before surgery and 3 months after surgery,corresponding dada of healing time,ankle brachial index (ABI) and Michigan neuropathy sensation instruments (MNSI) were closely observed in the patients of the 2 groups.Results The patients of the two groups all had medical followed-ups for an average of 9.3 months.In the control group,the wound of one patient with Wagner Ⅴ failed to heal and in the end had amputation,but the wound of the remaining patients all healed,with an average healing time of (39.52 ± 8.43) days.The wound of the treatment group all healed,with an average healing time of (31.74 ± 6.12) days.Three months after surgery,the ABI level of the treatment group increased from pre-surgical level 0.56 ± 0.18 to post-surgical level 0.94 ± O.16,and the MNSI level decreased from pre-surgical level 5.92 ± 1.13 to post-surgical level 4.11 ± 1.25.Three months after surgery,the ABI level of the control group increased from pre-surgical level 0.54 ±0.21 to post-surgical level 0.81 ±0.20,and the MNSI level decreased from pre-surgical level 5.74 ± 1.26 to post-surgical level 4.98 ± 1.34.Statistical significance could be noted in wound healing time when comparisons were made between the two groups.Three months after surgery,statistical significance could also be found in ABI and MNSI levels as compared with those detected before surgery.The treatment group was superior to the control group,also with statistical significance.Conclusions Lateral tibia bone removal micro vessel regeneration could be used to reconstruct vasoganglion below lower leg and promote surrounding vessel and nerve function recovery.Combined with HBO therapy,it could promote wound surface healing,with the advantages of simple surgery,accurate curative effect,high success rate of limb-salvage.For this reason,it was one of the ideal methods for the treatment of diabetic foot ulcer.
4.The mechanism of tumor necrosis factor-alpha participating in the osteoporosis of MRL/lpr mice by inhibiting osteoblast differentitation of BMMSCs in vivo
Dongming SONG ; Ting CUI ; Yingying QIU ; Jinbin RUI ; Xiaoming FEI ; Xinxin XU ; Jing LI ; Yu TANG
Chinese Journal of Rheumatology 2015;(6):364-368
Objective To investigate the mechanism of tumor necrosis factor-α (TNF)-α inhibiting osteo blastdifferentiation of mesenchymal stem cells (BMMSCs) in the pathogenesis of osteoporosis in the mouse model of systemic lupus erythematosus (MRL/lpr). Methods The femurs of MRL / lpr and C3He/HeJ mice were isolated, the bone structure were examined by hematoxylin-eosin (HE) staining. The proteins of TNF-α, NF-κB P50, bone morphogenetic protein -2 (BMP-2) and PSmad1/5/8 were measured by immunohistochemical stain. Bone marrow mesenchymal stem cells (BMMSCs) were isolated. After BMMSCs grew on the cover slips, the proteins on top of it were evaluated by immunohistochemistry stain. Moreover, the alkaline phosphatase (ALP) staining was employed for the measurement of the early osteogenic differentiation. BMMSCs together with hydroxyapatite were embedded subcutaneously in the nude mice and eight weeks later, the ectopic bone formation was evaluated. The recombinant human tumor necrosis factor receptor type Ⅱantibody fusion protein (etanercept) or normal saline was subcutaneous injected to the mice with lupus. After four weeks, the expression of these proteins was observed and the ectopic bone formation was investigated. Image-Pro plus 6.0 software was employed for imagine analysis, and Studentˊs t-test was used to test the differences between 2 independent groups. Results MRL/lpr mice showed decreased volume of cortex and the percentage of cortex to the volume of bone of MRL/lpr mice was significantly lower compared to control groups and with C3He/HeJ mice (13.96±0.25 vs 23.61±0.71, n=3, P<0.01). The protein levels of both TNF-αand NF-κB P50 on the femur of MRL/lprl mice were higher than those of the control group (0.643±0.051 vs 0.405±0.022, 0.917±0.023 vs 0.650±0.032, n=3, P<0.01). The expressions of BMP-2 on the femur of MRL/lpr mice were lower than those of the C3He/HeJ mice (0.52 ±0.03 vs 0.72 ±0.03, n=3, P<0.01). There was no difference in the expression of PSmad1/5/8 on the femur between the two groups by immunohistochemistry detection (1.264 ±0.021 vs 1.301± 0.044, n=3, P>0.05). The expressions of TNF-α and NF-κB P50 in BMMSCs of MRL/lprl mice were higher than those of the C3He/HeJ (0.184±0.021 vs 0.136±0.013, 0.132±0.021 vs 0.097± 0.014, n=3, P<0.01), while BMP-2 and PSmad were lower than those of the control group (0.128±0.013 vs 0.216±0.221, 0.115±0.023 vs 0.196±0.034, n=3, P<0.01). After 7 days of BMP-2 stimulation, the activities of ALP of BMMSCs from MRL/lprl mice were reduced detected by ALP staining and the osteoblast differentiation of these cells were decreased than BMMSCs from the control mice by HE and Masson staining. The percentage of the cortex to the volume of bone of the etanercept injection MRL/lpr mice was higher than that of the control group (21.8±1.0 vs 14.3 ±0.6, n=3, P<0.01). Moreover, the proteins of TNF-α and NF-κB P50 on the femurs of such injected mice were lower than those of the control group (0.540±0.024 vs 0.682±0.031, 0.857±0.023 vs 1.098±0.044, n=3, P<0.05), while the expressions of BMP-2 were higher than the control group (0.99±0.04 vs 0.85±0.04, n=3, P<0.05). There was no difference in the PSmad1/5/8 expression on the bone of the two group of lupus mice (0.88 ±0.08 vs 0.84 ±0.04, n=3, P>0.05). The ectopic bone formation of BMMSCs of the etanercept injected MRL/lpr mice was higher than that of the normal saline injected mice, however, it was lower than that of the C3He/HeJ mice. Conclusion TNF-α inhibits osteoblast differentiation of mesenchymal stem cells by depressing Smad signaling which may contribute to the osteoporosis of the lupus mice.
5.Correction of the complex rigid talipes equinovarus deformities with the Ilizarov technique
Feng CHANG ; Bin CHEN ; Zhizhen JING ; Gang GAO ; Lijun LI ; Jinbin WEI ; Dean QIN ; Xiaojian WANG ; Jianping YU ; Jiefu SONG ; Yunxing SU
Chinese Journal of Orthopaedics 2012;32(3):222-228
Objective To evaluate the clinical results of the Ilizarov technique for the treatment of the complex rigid talipes equinovarus deformities.Methods From July 2005 to July 2011,28 patients (41 feet) with rigid talipes equinovarus deformities which had been corrected with the Ilizarov technique were retrospectively analyzed,including 18 males (26 feet) and 10 females (15 feet) with an average age of 15.3 years.According to the classification system proposed by Diméglio,31 feet were categorized as grade Ⅲ,and 10 as grade Ⅳ.We performed corrections with a soft tissue release in 23 feet,and with a limited osteotomy in 18,and then a Ilizarov external fixator was applied.Anteroposterior and lateral X-rays were taken to compare the pre and postoperative data in terms of the angle of plantarflexion and dorsiflexion,the range of motion of the ankle joint,radiological measurements of the talocalcaneal angle.Results All the 28 patients achieved an outpatient follow-up,with an average of 25 months.All patients achieved a plantigrade foot with an almost normal appearance as the fixator was removed after applied for an average of 5.1 months (range,2-14).At the preoperative and final follow-up respectively,the angle of dorsiflexion of the foot was -45.0°±12.0° and 9.5°±5.5°,the angle of plantarflexion was 67.0°±14.0° and 45.5°±7.8°,talocalcaneal angle was 6.5°±4.5° and 22.5°±5.5° in anteroposterior radiograph and 5.5°±11.0° and 40.6°±8.5° in lateral radiograph.Spastic ischemia occurred in one foot and relieved by a slower distraction rate.Wire-hole infections occurred in 5 feet and treated by dressing changs,wire tract altering and antibiotic therapy,finally the infections were controlled.Deformity relapsed in one foot three months after the device was removed,then corrected with an additional fixator application and has not recurred till the final follow-up.Toe contracture and residual deformity occurred in 5 feet and 3 feet,respectively.Conclusion The Ilizarov technique is an effective method for correction of complex rigid talipes equinovarus deformities,with which the appearance and function of the foot could be kept as much as possible,and without impact on food development.

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