BACKGROUND: It has been reported that Angiotensin Ⅱ (Ang Ⅱ) is related to occurrence and development of dermatofibrosis; however, less is explored about the expression and effect of AT1 and AT2 receptors in the fibroblasts of human hypertrophic scar.OBJECTIVE: To observe the expression of Ang Ⅱ type 1 (AT1) and type 2 (AT2) receptors in human hypertrophic scars, and explore their effects on collagen synthesis of fibroblasts.DESIGN, TIME AND SETTING: Randomized control experiment was performed at the Experimental Center, Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA between August 2006 and November 2007. PARTICIPANTS: Samples of hypertrophic scare were taken from 18 patients (10 males and 8 females, 19-47 years Old). Seven specimens of normal skin served as control. All of the specimens collected were divided into two parts, one part for immunohistochemical staining after fixated by 4% paraformaldehyde, the other part for culturing fibroblasts.METHODS: The expression of both AT1 and AT2 receptors in fibroblasts of hypertrophic scare was detected with immunohistochemical staining and radioligand receptor binding assay. Collagen synthesis was examined in cultured fibroblasts of hypertrophic scars by measuring [3H]-proline incorporation into collagenous proteins.MAIN OUTCOME MEASURES: The expression of both AT1 and AT2 receptors in human hypertrophic scars; the [3H]-proline incorporation value in cultured fibroblasts.RESULTS: Positive staining signals of both AT1 and AT2 receptors were found in fibroblasts of hypertrophic scars. Similar results were also observed in cultured fibroblasts of hypertrophic scars, expression level of AT1 and AT2 receptors were (10.69±2.15) fmol/106 cells and (4.9±1.05) fmol/106cells, respectively. In cultured fibreblasts, Ang Ⅱ stimulation significantly increased collagen synthesis, which was inhibited by valsartan, an AT1 receptor blocker, but augmented by PD123319, an AT2 receptor antagonist.CONCLUSION: Both AT1 and AT2 receptors were expressee in the fibreblasts of hypertrophic scars, and Ang Ⅱ regulates collagen synthesis in hypertrophic scar fibroblasts through a negative cross-talk between AT1 and AT2 receptors, which might contribute, at least partly to formation and maturation of human hypertrophic scars.

Obje:ctive To establish an optimized method to isolate, culture and identify human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro and induce their osteogenic and adipogenic differentiation. Methods The hUCMSCs were isolated from human umbilical cord by digestion with collagenase. After serial subcultivation in vitro, the stem cells were passaged. Morphologic appearance of hUCMSCs was observed under an optical microscope and atomic force microscope. The proliferation rate was measured by MTT assay. Cell cycle and surface antigens were measured by flow cytometry. The osteogenic and adipogenic differentiation was tested and evaluated by specific staining methods. Results The isolation of hUCMSCs by digestion with collagenase was efficient. After seeded for 24 hours, the adherent cells showed spindle shape and fibroblast cell-like shape and the size of hUCMSCs was homogeneous. The similar growth curves of passage 3 and 7 exhibited a great potential for proliferation. Flow cytometry analysis revealed that CD29, CD44 and CD105 were highly expressed on the surface of passages 3 cells, but the expression was negative for CD34, CD45 and HLA-DR. After culture in inducing medium, the cells were successfully induced into osteogenic and adipogenic lineages. These cells were highly positive for alkaline phosphate staining and also showed mineralization presented with von kossa staining after 4 weeks' culture induction of osteogenic differentiation. Furthermore, liquid vacuoles were detected by oil red O staining after 3 weeks' culture induction of adipogenic differentiation. Conclusion An in vitro method for isolation and purification of hUCMSCs from human umbilical cord has been established. The cultured cells were composed of only undifferentiated cells and their biological properties were stable. The hUCMSCs are expected to be a new type of stem cells of tissue engineering.

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 .

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

Objective: To analyze the expression of tight junction protein 3(claudin-3) in colorectal cancer and its relationship with the development, metastasis and prognosis of colorectal cancer. Methods: From February 2013 to February 2015, 78 patients with colorectal cancer operated in the People's Hospital of Sanmen County were selected in this study.The tissues of colorectal cancer and adjacent tissues were collected and claudin-3 expression was detected.The relationship between claudin-3 and clinicopathological features of colorectal cancer was analyzed. Results: The positive rate of claudin-3 in cancer tissues(83.33%) was higher than that in paracancerous tissues(48.72%), and the difference was statistically significant(χ²=20.832, P<0.01). The positive rate of claudin-3 in the poorly differentiated group(100.00%) was significantly higher than that in the well-differentiated group(85.71%) and the well-differentiated group(52.63%)(χ²=19.209, P<0.01). The claudin-3 positive rate with lymphatic metastasis(97.30%) was higher than that without lymphatic metastasis(70.73%), and the difference was statistically significant(χ²=9.882, P<0.01). The claudin-3 positive rate in patients with less than 3 years of survival (91.11%) was higher than that in patients with more than 3 years of survival(72.73%)(χ²=4.633, P<0.05). Conclusion The expression of claudin-3 is related to the occurrence, development and lymphatic metastasis of colorectal cancer, and can predict the prognosis of the patients.

OBJECTIVETo evaluate the clinical effect of different surgical approaches for treating cervical ossification of the posterior longitudinal ligament (OPLL) with spinal cord signal change.

METHODSThirty-eight patients with OPLL with spinal cord signal change were treated from January 2005 to January 2011. Surgical removal via an anterior approach or partial decompression was performed in 10 cases (group A), posterior approach open-door laminoplasty with decompression, bone grafting and internal fixation was performed in 12 cases (group B), and opening the cervical spinal meninges to relieve the pressure was performed in 16 cases (group C) on the basis of the procedures in group B. All the patients were followed up and the pre- and postoperative JOA scores, improvement ratio and inter-body implant fusion were evaluated. Imaging examinations including X-rays, CT and MRI were also performed pre- and postoperatively, and the surgical complications were recorded.

RESULTSAt 12 months postoperatively, the mean improvement rates in groups A, B, and C were 52.39%, 55.15%, and 60.32%, respectively, with the mean JOA scores of 13.54∓0.56, 13.56∓1.26, and 14.70∓1.41, respectively. The JOA scores and improvement rates significantly increased after the surgeries. One patient in group A became paraplegic after the operation with cerebrospinal fluid leakage, and one patient in group B and one in group C reported numbness of the upper limb. Group C showed a shorter postoperative recovery time without severe complications.

CONCLUSIONPosterior open-door laminoplasty, decompression, bone grafting and internal fixation can be an effective approach for treatment of cervical OPLL with spinal cord signal change and requires shorter rehabilitation time after the operation.

Aged ; Cervical Vertebrae ; pathology ; Decompression, Surgical ; methods ; Female ; Humans ; Male ; Middle Aged ; Ossification of Posterior Longitudinal Ligament ; pathology ; surgery ; Spinal Cord Compression ; etiology ; surgery ; Treatment Outcome

Objective: To investigate the safety and long-term efficacy of a new treatment, echocardiography-guided transthoracic laser ablation of the animal interventricular septum (IVS), for obstructive hypertrophic cardiomyopathy (HOCM). Methods: Ten healthy sheep were randomly divided into two groups: experimental group: sheath puncture with laser ablation (energy: 3 W, 1000 J), sham control group: sheath puncture only without laser ablation. Echocardiography and electrocardiogram (ECG) were recorded before operation, immediately after operation, and 1, 3 and 6-month after the operation. Left ventricular systolic and diastolic function, longitudinal strain, difference of time to peak between the ablation segment and the surrounding segments were analyzed. Blood samples were collected before and one hour after the operation to examine the serological results. Results: Immediately and 6 months after the operation, all animals survived with normal cardiac function. No severe complications such as cardiac tamponade or bundle branch block occurred. The Troponin I level was significantly elevated immediately after the operation(P<0.05). The thickness of the ablated IVS was significantly reduced 6 months after the operation compared with before the operation[(3.23±1.21)mm vs (8.53±0.44)mm, P<0.05]. M-mode echocardiography showed that the amplitude of movement of the ablated region of the experimental group 6 months after the operation was significantly decreased compared with the control group(P<0.05). Three dimensional strain analysis showed that for the experimental group, the longitudinal strain of the ablation segment was significantly reduced and the difference of time to peak was significantly delayed, compared with the control group(P<0.05). Conclusions Echocardiography-guided transthoracic laser ablation of IVS is a safe, effective, and minimally invasive method. It is capable to reduce the volume of IVS without influencing the cardiac function, which makes it a potential alternative for HOCM treatment.