OBJECTIVE: To evaluate the capacity and efficiency of human umbilical cord mesenchymal stem cells (HUCMSCs) to differentiate into neuron- like cells after induction with B27- supplemented serum- free medium. METHODS: HUCMSCs at passage 4 were cultured for 14 days with serum-containing medium (SCM) (group A), SCM supplemented with 20 ng/mL nerve growth factor (NGF) and 10 ng/mL basic fibroblast growth factor (bFGF) (group B), serum-free medium (SFM) (group C), or SFM supplemented with 20 ng/mL NGF and 10 ng/mL bFGF. The culture medium were changed every 3 days and the growth of the neurospheres was observed using an inverted microscope. The cell markers were analyzed with flow cytometry and the expressions of nestin, neuron- specific enolase (NSE), neurofilament heavy polypeptide (NEFH), and glial fibrillary acidic protein (GFAP) were quantified by quantitative real-time PCR (qRT-PCR) and Western blotting. RESULTS: Before induction, HUCMSCs expressed abundant mesenchymal stem cell surface markers including CD29 (99.5%), CD44 (49.6%) and CD105 (77.7%). Neuron-like cells were observed in the cultures on days 7, 10, and 14, and the cell differentiation was the best in group D, followed by groups C, B and A. In all the 4 groups, the cellular expressions of nestin and GFAP gradually lowered while those of NEFH and NSE increased progressively. The expressions of GFAP, NEFH, nestin and NSE were significantly different between group A and the other 3 groups ( < 0.001 or 0.05). CONCLUSIONS B27-supplemented SFM effectively induces the differentiation of HUCMSCs into neuron- like cells, and the supplementation with cytokines (NGF and bFGF) strongly promotes the cell differentiation.

Recurrent laryngeal nerve (RLN) injury can result in unilateral or bilateral vocal cords paralysis, thereby causing a series of complications, such as hoarseness and dyspnea. However, the repair of RLN remains a great challenge in current medicine. This study aimed to develop human umbilical mesenchymal stem cells (HuMSCs) and nerve growth factor (NGF)-loaded heparinized collagen scaffolds (HuMSCs/NGF HC-scaffolds) and evaluate their potential in the repair of RLN injury. HuMSCs/NGF HC-scaffolds were prepared through incorporating HuMSCs and NGF into heparinized collagen scaffolds that were prefabricated by freeze-drying in a template. The resulting scaffolds were characterized by FTIR, SEM, porosity, degradation in vitro, NGF release in vitro and bioactivity. A rabbit RLN injury model was constructed to appraise the performance of HuMSCs/NGF HC-scaffolds for nerve injury repair. Electrophysiology, histomorphology and diagnostic proteins expression for treated nerves were checked after application of various scaffolds. The results showed that the composite scaffolds with HuMSCs and NGF were rather helpful for the repair of broken RLN. The RLN treated with HuMSCs/NGF HC-scaffolds for 8 weeks produced a relatively normal electromyogram, and the levels of calcium-binding protein S100, neurofilament and AchE pertinent to nerve were found to be close to the normal ones but higher than those resulted from other scaffolds. Taken together, HuMSCs/NGF HC-scaffolds exhibited a high score on the nerve injury repair and may be valuable for the remedy of RLN injury.
Collagen* ; Dyspnea ; Electrophysiology ; Heparin* ; Hoarseness ; Humans* ; In Vitro Techniques ; Intermediate Filaments ; Mesenchymal Stromal Cells* ; Nerve Growth Factor* ; Paralysis ; Porosity ; Recurrent Laryngeal Nerve Injuries* ; Recurrent Laryngeal Nerve* ; Spectroscopy, Fourier Transform Infrared ; Umbilical Cord* ; Vocal Cords

Objective To evaluate the efficacy of minimally invasive percutaneous plate osteosynthesis (MIPPO) in the treatment of Neer three-part fractures of the proximal humerus in young adults. Methods A retrospective analysis was performed of the 46 patients aged < 65 years with Neer three-part fracture of the proximal humerus from March 2010 to December 2016. MIPPO with locking proximal humerus plate ( LPHP ) was used in 23 of them who were 12 men and 11 women with an average age of 41. 6 ± 1. 2 years; open reduction and internal fixation ( ORIF ) with LPHP was used in the other 23 patients who were 14 men and 9 women with an average age of 42. 2 ± 1. 6 years. The 2 groups were compared in terms of operation time, intraoperative bleed-ing, fracture healing time and shoulder function by Neer scoring at the last follow-up. Results The average follow-up ( 13. 4 ± 1. 2 months ) for the MIPPO groups was not statistically different from that for the ORIF group ( 14. 2 ± 2. 4 months ) ( P > 0. 05 ) . The MIPPO group reported significantly shorter operation time ( 105 ± 15 min ) , significantly less intraoperative bleeding ( 140 ± 50 mL ) , significantly shorter fracture healing time ( 4. 2 ± 0. 6 months ) , and significantly higher shoulder Neer scores ( 88. 6 ± 3. 4 ) than the ORIF group ( 120 ± 20 min, 320 ± 40 mL, 5. 4 ± 1. 2 months, and 81. 6 ± 2. 2, respectively ) ( P <0. 05 ) . The complication rate ( 4. 3％, 1/23 ) for the MIPPO group was not significantly different from that for the ORIF group ( 17. 4％, 4/23 ) ( P >0. 05 ) . Conclusion MIPPO with LPHP may be obviously advantageous over ORIF with LPHP in the treatment of proximal humeral fractures in young adults.

OBJECTIVETo investigate the effect of ganoderic acid A (GA-A) on the biological behaviors of human osteosarcoma cells in vitro.

METHODSMG63 and HOS cells were treated with 0.1, 0.25, and 0.5 mmol/L GA-A, and the changes in cell proliferation, apoptosis and migration were evaluated using MTT assay, flow cytometry, and Transwell assay, respectively. The expressions of STAT3, p38, and NF-κB1 in the cells were analyzed by Western blotting.

RESULTSGA-A effectively inhibited the proliferation of human osteosarcoma HOS and MG-63 cells in a dose-dependent manner, and induced obvious cell apoptosis in both cells. Treatment with 0.5 mmol/L GA-A also resulted in significant inhibition of the invasion of both cells. The results of Western blotting showed that GA-A down-regulated the expression level of phosphorylated STAT3 and increased the phosphorylation level of p38 and NF-κB1 expression in both cells.

CONCLUSIONGA-A can induce proliferation inhibition, apoptosis and suppression of invasion in human osteosarcoma HOS and MG-63 cells.

Apoptosis ; drug effects ; Bone Neoplasms ; pathology ; Cell Line, Tumor ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Heptanoic Acids ; pharmacology ; Humans ; Lanosterol ; analogs & derivatives ; pharmacology ; NF-kappa B p50 Subunit ; metabolism ; Osteosarcoma ; pathology ; Phosphorylation ; STAT3 Transcription Factor ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

AIM:To explore an ideal method to induce the differen-tiation of human umbilical cord mesenchy-mal stem cells (hUCMSCs) into neuron-like cells and to provide some evidence for the transplantation of hUCMSCs for spi-nal cord injury .METHODS:The hUCMSCs were isolated from human umbilical cord digested with collagenase Ⅱ.The hUCMSCs was verified by flow cytometry analysis .The passage 5 cells were randomly divided into 4 groups.The differentiation of hUCMSCs was induced by bFGF in group A , bFGF and BDNF in group B, or BHA, bFGF and BDNF in group C, while the cells in group D served as a control group cultured with DMEM-F12 and 10%FBS.Two weeks later , the expression of nestin , neurofilament protein H ( NEFH) and glial fibrillary acidic protein ( GFAP) was detected by real-time PCR and immunocytochemistry .The morphological changes of cells were observed under an atomic force microscope . RESULTS:Mesenchymal stem cells were isolated and cultured from human umbilical cord by enzyme digestion .hUCMSCs expressed CD29, CD44 and CD105, but no CD34, CD45 or HLA-DR.After cultured with inducing medium for 2 weeks, the cells were successfully induced into neuron-like cells.The appearance of the cells had great change .The induced hUC-MSCs developed round cell bodies with multiple neurite-like extensions observed under an atomic force microscope .The re-sult of real-time PCR showed that nestin was positive in A , B and C groups , and NEFH was positive in A and B groups , but GFAP was negative in 4 groups.The difference of nestin and NEFH expression among the induced groups was signifi -cant (P<0.05).CONCLUSION:Mesenchymal stem cells were isolated and cultured from human umbilical cord by en-zyme digestion in vitro, and all the hUCMACs presented stable biological properties .Moreover, hUCMSCs were induced to differentiate into neuron-like cells in vitro via bFGF combined with BDNF .

Objective To investigate the effect of ganoderic acid A (GA-A) on the biological behaviors of human osteosarcoma cells in vitro. Methods MG63 and HOS cells were treated with 0.1, 0.25, and 0.5 mmol/L GA-A, and the changes in cell proliferation, apoptosis and migration were evaluated using MTT assay, flow cytometry, and Transwell assay, respectively. The expressions of STAT3, p38, and NF-κB1 in the cells were analyzed by Western blotting. Results GA-A effectively inhibited the proliferation of human osteosarcoma HOS and MG-63 cells in a dose-dependent manner, and induced obvious cell apoptosis in both cells. Treatment with 0.5 mmol/L GA-A also resulted in significant inhibition of the invasion of both cells. The results of Western blotting showed that GA-A down-regulated the expression level of phosphorylated STAT3 and increased the phosphorylation level of p38 and NF-κB1 expression in both cells. Conclusion GA-A can induce proliferation inhibition, apoptosis and suppression of invasion in human osteosarcoma HOS and MG-63 cells.

Objective To investigate the effect of ganoderic acid A (GA-A) on the biological behaviors of human osteosarcoma cells in vitro. Methods MG63 and HOS cells were treated with 0.1, 0.25, and 0.5 mmol/L GA-A, and the changes in cell proliferation, apoptosis and migration were evaluated using MTT assay, flow cytometry, and Transwell assay, respectively. The expressions of STAT3, p38, and NF-κB1 in the cells were analyzed by Western blotting. Results GA-A effectively inhibited the proliferation of human osteosarcoma HOS and MG-63 cells in a dose-dependent manner, and induced obvious cell apoptosis in both cells. Treatment with 0.5 mmol/L GA-A also resulted in significant inhibition of the invasion of both cells. The results of Western blotting showed that GA-A down-regulated the expression level of phosphorylated STAT3 and increased the phosphorylation level of p38 and NF-κB1 expression in both cells. Conclusion GA-A can induce proliferation inhibition, apoptosis and suppression of invasion in human osteosarcoma HOS and MG-63 cells.

OBJECTIVETo evaluate the safety and efficacy of Endostar combined with chemotherapy in the treatment of end-stage colorectal cancer.

METHODSs The relevant randomized controlled trials were retrieved from the electronic databases of Cochrane library, PubMed, EMbase, CNKI, CBM, VIP and Chinese Medical Association. The retrieval time limit was from the database construction to January 2013. The data were extracted from eligible studies assessed for methodological quality according to Cochrane handbook for systematic reviews and analyzed using RevMan 5.2 software.

RESULTSFive randomized controlled trials involving 220 cases were included for meta-analysis. The results showed that Endostar combined with chemotherapy had an overall advantage over chemotherapy alone in terms of complete response rate (10.91% vs 2.73% RR=4.08, 95% CI: 1.19-13.95, P=0.02), partial response rate (48.18% vs 30.91% RR=2.18, 95% CI: 1.23-3.87, P=0.007), progressive disease (15.45% vs 41.82% RR=0.25, 95% CI: 0.13-0.47, P<0.0001), and the response rate (60.00% vs 33.64% RR=3.23, 95% CI: 1.79-5.81, P<0.0001). Clinical benefit response(82.73% vs 55.45% RR=4.30,95% CI:1.19-13.95, P<0.0001). The main adverse reactions included nausea, vomiting, constipation, palpitation, and electrocardiogram changes.

CONCLUSIONEndostar combined with chemotherapy is effective for advanced colorectal cancer and can be used as a routine treatment.

Antineoplastic Agents ; administration & dosage ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Colorectal Neoplasms ; drug therapy ; Endostatins ; administration & dosage ; Humans ; Randomized Controlled Trials as Topic

Objective To evaluate the safety and efficacy of Endostar combined with chemotherapy in the treatment of end-stage colorectal cancer. Methods The relevant randomized controlled trials were retrieved from the electronic databases of Cochrane library, PubMed, EMbase, CNKI, CBM, VIP and Chinese Medical Association. The retrieval time limit was from the database construction to January 2013. The data were extracted from eligible studies assessed for methodological quality according to Cochrane handbook for systematic reviews and analyzed using RevMan 5.2 software. Results Five randomized controlled trials involving 220 cases were included for meta-analysis. The results showed that Endostar combined with chemotherapy had an overall advantage over chemotherapy alone in terms of complete response rate (10.91%vs 2.73%RR=4.08, 95%CI:1.19-13.95, P=0.02), partial response rate (48.18% vs 30.91% RR=2.18, 95% CI: 1.23-3.87, P=0.007), progressive disease (15.45% vs 41.82% RR=0.25, 95% CI: 0.13-0.47, P<0.0001), and the response rate (60.00% vs 33.64% RR=3.23, 95% CI: 1.79-5.81, P<0.0001). Clinical benefit response(82.73%vs 55.45%RR=4.30,95%CI:1.19-13.95, P<0.0001). The main adverse reactions included nausea, vomiting, constipation, palpitation, and electrocardiogram changes. Conclusion Endostar combined with chemotherapy is effective for advanced colorectal cancer and can be used as a routine treatment.

Objective To evaluate the safety and efficacy of Endostar combined with chemotherapy in the treatment of end-stage colorectal cancer. Methods The relevant randomized controlled trials were retrieved from the electronic databases of Cochrane library, PubMed, EMbase, CNKI, CBM, VIP and Chinese Medical Association. The retrieval time limit was from the database construction to January 2013. The data were extracted from eligible studies assessed for methodological quality according to Cochrane handbook for systematic reviews and analyzed using RevMan 5.2 software. Results Five randomized controlled trials involving 220 cases were included for meta-analysis. The results showed that Endostar combined with chemotherapy had an overall advantage over chemotherapy alone in terms of complete response rate (10.91%vs 2.73%RR=4.08, 95%CI:1.19-13.95, P=0.02), partial response rate (48.18% vs 30.91% RR=2.18, 95% CI: 1.23-3.87, P=0.007), progressive disease (15.45% vs 41.82% RR=0.25, 95% CI: 0.13-0.47, P<0.0001), and the response rate (60.00% vs 33.64% RR=3.23, 95% CI: 1.79-5.81, P<0.0001). Clinical benefit response(82.73%vs 55.45%RR=4.30,95%CI:1.19-13.95, P<0.0001). The main adverse reactions included nausea, vomiting, constipation, palpitation, and electrocardiogram changes. Conclusion Endostar combined with chemotherapy is effective for advanced colorectal cancer and can be used as a routine treatment.