ObjectiveTo illustrate the role of epidermal growth factor (EGF) secreted by astrocytes in the process of tacrolimus (FK506) in promoting neurite outgrowth. MethodsThe spinal cord astrocytes and neuronal cells were isolated respectively from 2-day-old Sprague Dawley (SD) rats and 15-day SD pregnant rats, and cultured in vitro and identified by immunofluorescence staining. The spinal cord astrocytes were cultured with 20 μmol/L FK506 medium in the experimental group, and with FK506 free medium in the control group. The supernatant was collected after 24 hours for preparing conditioned medium, and astrocytes were collected. EGF proteins in the conditioned medium were detected with ELISA, and EGF gene expressions of astrocytes were detected with real-time quantitative PCR (RT-qPCR). The spinal cord neurons were cultured respectively with conditioned medium from the experimental group (FK506-CM) and the control group (C-CM) in group A and group B, also with neutralized C-CM and neutralized FK506-CM with anti-EGF neutralizing antibodies in group C and group D. Both the total neurite length and the longest neurite length were measured and compared among groups. ResultsBoth astrocytes and neurons were confirmed by immunofluorescence staining. The EGF content of experimental group (0.241±0.044) was significantly higher than that of the control group (0.166±0.014) (t=3.93, P=0.01); EGF gene expression of the experimental group (1.12±0.25) was significantly higher than that of the control group (0.46±0.11) (t=5.78, P=0.00). The neurite length measurement displayed that the total neurite length and the longest neurite length of groups C and D were significantly shorter than those of groups A and B (P<0.05). Both the total and longest neurite length of group A were significantly longer than those of group B (P<0.05), but no significant difference was shown between groups C and D (P>0.05). ConclusionThe EGF secreted by spinal cord astrocytes can promote the neurite outgrowth. So spinal cord astrocytes can be used as an important intermediary target of FK506 to promote the recovery of neurological function.
ObjectiveTo summarize the surgical strategy on treating mitral desease patient associated with hypertrophic obstructive cadiomyopathy (HOCM). MethodsWe retrospectively analyzed the clinical data of 17 patients with HOCM underwent surgical treatment from November 2003 to May 2015 year. There were 10 males and 7 females with a mean age of 42.2±15.5 years ranging from 7-62 years. There were 16 patients underwent modified Morrow procedure and 1 patient underwent modified Konno procedure to relieve the obstruction of left ventricular outflow tract. And different surgical treatment of mitral valve disease was implemented depending on the severity of regurgitation and under monitoring of transesophageal echocardiography. About 2 weeks after the surgery, we performed transthoracic echocardiography to evaluate the effect of operation. ResultsNo hospital death occurred and the surgery obviously improved the symptom and cardiac function in all cases. After surgery, echocardiography revealed that the mean thickness of the ventricular septum statistically decreased (P < 0.0001), the systolic anterior motion disappeared, the outflow track pressure of left ventricle statistically decreased (P < 0.0001), and the peak flow rate of left ventricle statistically decreased. However, there was no statistical difference in the change of the left ventricular ejection fraction(P=0.083). Nine patients with no mitral regurgitation (MR) or mild MR only underwent the unblock of the left ventricular outflow track, the MR decreased to mild or disappeared. Four patients with moderate or severe MR underwent mitral valve repair, and the MR decrease to mild or disappear. There were no complications occurred regarding to prosthesis implantation over the 4 patients underwent mitral valve replacement for infective endocarditis or other causes. ConclusionFor the HOCM patients with mild MR, the unblock of the left ventricular outflow track alone can effectively improve the MR. For those combined with moderate or severe MR, we should choose mitral valve repair or replacement based on individual situation of patient.
ObjectiveTo compare the anti-apoptotic potency of human mesenchymal stem cells (hMSCs) derived from patients with cyanotic congenital heart diseases (C-CHD) or acyanotic congenital heart diseases (A-CHD) in vitro and explore the possible mechanism. MethodshMSCs were isolated from patients with cyanotic (Group C) or acyanotic (Group A) congenital heart diseases and cultured in a hypoxic incubator (1% O2, 5% CO2, 94% N2) in vitro. The anti-apoptotic potency of the hMSCs was assayed by the Annexin V-FITC/PI double labeled flow cytometry. The content of B-cell lymphoma-2 (Bcl-2), Bax and caspase-3 in both groups was determined by Western blot. ResultsFlow cytometry results revealed that hMSCs from C-CHD patients presented higher level of resistance to ischemia-and anoxia-induced apoptosis with lower overall (P<0.05) and early apoptosis ratio (P<0.01). Further Western blot examination identified that C-CHD-derived hMSCs produced more Bcl-2 (P<0.05) but less Bax (P<0.05) and caspase-3 (P<0.05) in comparison to their A-CHD-derived ones. ConclusionC-CHD-derived hMSCs presented the superiority for the anti-apoptotic potential, and the possible mechanism is the favorable change of Bcl-2, Bax and caspase-3 induced by the natural hypoxic and anoxic precondition.