Development and optimization of an effective method for evaluating habituation learning behavior in larval zebrafish.

BACKGROUND: Habituation learning is a simple and conserved behavior in all organisms which could be induced by repeated stimuli. However, no standard and universal methods for training and evaluating the habituation learning behavior in larval zebrafish were available. NEW METHOD: This study aims to establish effective training and detection protocols for habituation learning behavior in larval zebrafish by using the ViewPoint system. For this purpose, the detection threshold of velocity-a parameter for distinguishing the escape reaction and the spontaneous motion, the detection sensitivity-a parameter for determining the size of the identified object, the number of stimuli, and the age of larvae were optimized to obtain the best performance. RESULTS: In this study, the optimized parameters were as follows: the detection threshold of velocity at 13, the luminous intensity at 8 %, the detection sensitivity at 32, the number of stimuli at 150, and the age of larvae at 6 dpf. Furthermore, we validated the utility of the established protocol by showing a consistent memory impairment induced by cycloheximide (CHX). COMPARISON WITH EXISTING METHOD: A similar method was reported previously. However, the equipment used in those assays, including the hardware and software, were neither standard nor universal, which might impede the extensive application of the habituation learning assays. Here, we developed an alternative method for studying the habituation learning behavior in larval zebrafish using the ViewPoint system. CONCLUSIONS: Our study provided an alternative method for studying the habituation learning behavior in larval zebrafish.

Tetramethylpyrazine/Ligustrazine Can Improve the Survival Rate of Adipose-Derived Stem Cell Transplantation.

BACKGROUND: We aimed to study the effect of tetramethylpyrazine/ligustrazine (TMP) on the survival rate of adipose-derived stem cell (ADSC) transplantation. METHODS: Human ADSCs were cultured and subcultured. Detection of cell growth was performed using CCK8 kit. A total of 12 BALB/c nude mice including 4 groups (group 1: ADSC-hyaluronic acid [HA] + phosphate-buffered saline [PBS]; group 2: ADSC-HA + TMP injection; group 3: ADSC-HA + TMP soaking; group 4: ADSC-HA + TMP injection + TMP soaking) were used for the animal experiments. Furthermore, hematoxylin-eosin staining and oil red O staining were conducted for the frozen section from the animal tissues. Besides, RNA was extracted from the samples, and reverse transcriptase-polymerase chain reaction experiment was performed. At last, samples were detected using transmission electron microscope. RESULTS: Tetramethylpyrazine/ligustrazine resulted in significant increase for cell proliferation on the seventh day. It was easier to promote the growth of adipose cell after injection and soaking of TMP. Hematoxylin-eosin staining and oil red O stain showed that injection and soaking of TMP could significantly increase the size of fat. The relative expression of peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein α, and Alu in group 4 was significantly higher than that in other groups. CONCLUSIONS: Tetramethylpyrazine/ligustrazine can improve the survival rate of ADSC transplantation. The possible mechanisms of it may be that TMP induces the expression of transcription factor associated with fat formation including peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein α, and Alu and then promotes the growth of adipose cell.

Staged reconstructive treatment for extensive irregular cicatricial alopecia after burn.

For extensive irregular cicatricial alopecia after burn, effective and pleasing restoration of hair-bearing scalp remains challenging. In this article, the authors presented staged reconstructive treatment for extensive irregular cicatricial alopecia with the goal to achieve better and reliable results. A retrospective review of staged reconstructive treatment performed in 16 patients with extensive irregular cicatricial alopecia after burn was conducted. In stage 1, final flaps were designed at 1st. Tissue expanders were placed into the subgaleal plane and serially inflated with normal saline. In stage 2, scarring tissues were excised and the expanded hair-bearing flaps were advanced to the defect. Hair grafts were harvested from excessive hair-bearing scalps excised from the flaps and replanted. For patients with less satisfactory results, stage 3 was performed by hair transplantation. Cicatricial area, follicular unit density, survival rate of hair grafts, and patients' satisfaction were measured before and after each stage. Thirteen patients received 3-stage treatment, and 3 received 2-stage treatment. Significant improvements in aesthetics and patient satisfaction were achieved in all the patients. No flap necrosis, implant exposure or hematoma was observed. Ideal, aesthetic, and reliable results could be obtained using staged reconstructive treatment for patients with extensive irregular cicatricial alopecia after burn.

[Analysis of outcome and prognostic factors of anterior approach for two-level cervical spondylotic myelopathy].

OBJECTIVE: To investigate the efficacy and possibly prognositic factors of anterior approach on two-level cervical spondylotic myelopathy. METHODS: A retrospective review was performed on 44 cases of two-level cervical spondylotic mydopathy from Jun. 2007 to Sep. 2009. Among the patients, 24 cases were male and 20 cases female, with an average age of (60.072 +/- 10.77) years (ranged from 39 to 80 years). The affected segments ranged from C(3.4)-C(6.7). Improvements of cervical curvature and segmental height, preoperative sagittal diameter of the spinal cord at the site of maximal compression and signal intensity changes on T2WI were respectively measured. Function of nerves was assessed according to Japanese orthopaedic association system (JOA:17 score) before and after surgery. Operation by anterior approach including: anterior corpectomy and titanium cage fusion with internal fixation, anterior corpectomy body and auto iliac bone fusion with internal fixation. Statistical analysis was made on the correlation between JOA recovery rate and prognostic factors. RESULTS: Improvements of cervical curvature was -9.1 degrees to 16.6 degrees with the mean of (1.30 +/- 5.77) degrees and improvements of segmental height was -0.3 to 12.3 mm with the mean of (4.23 +/- 3.08) mm. Sagittal diameter of the spinal cord at the site of maximal compression was 1.6 to 7.2 mm with the mean of (4.01+/- 1.25) mm. T2WI with high signal changes was in 29 cases, no change in 15 cases. Bleeding amount was 50 to 700 ml with the mean of (242.05 +/- 148.22) ml. Operative time was 90 to 250 min with the mean of (153.75 +/- 34.54) min. All patients were followed up from 6 to 31 months with an average of (17.18 +/- 7.41) months. The mean JOA score preoperatively was (12.73 +/- 2.23); at the final follow-up, the JOA score was(15.09 +/- 1.91); and the recovery rate was (60.01 +/- 26.98)%. According to standard of JOA scoring, 16 cases obtained excellent result, 12 good, 15 fair and 1 poor. The effect of anterior approach had correlations with age, time of course, preoperative JOA score, sagittal diameter of the spinal cord at the site of maximal compression and signal intensity changes on T2WI, but had no correlations with operation time, bleeding amount and improvements of cervical curvature and segmental height. CONCLUSION: Two-level cervical spondylotic myelopathy can achieve good effect through anterior approach. The extent of the spinal cord compression may be a reliable and direct factor to judge effect.