Adipose tissue is a readily available source of adult stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Peptide hydrogel is a novel biomaterial which provides three-dimensional microenvironments for a variety of cells for tissue grafting. In this study, adipose-derived stem cells (ADSCs) were isolated from rats, seeded into the peptide hydrogel polymer scaffolds and cultured in Neurobasal (NB) media supplemented with B27, basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). Ten days after the culture, some cells were expanded into clonal populations in which the expression of both Nestin and Brdu was detected but only Brdu expression was detected in the cells that were not expanded into clonal populations. Our results suggested that ADSCs in peptide hydrogel polymer scaffolds can be induced to differentiate into cells capable of expressing the neuron-associated markers, self-renewal and self-propagation.

BACKGROUND:Kidney-tonifying herbs have been reported to promote tendon-bone healing in rabbits after anterior cruciate ligament reconstruction (ACLR). OBJECTIVE:To verify the effects of kidney-tonifying herbs on tendon-bone healing after ACLR. METHODS:Seventy patients with anterior cruciate ligament injury undergoing ACLR were randomly assigned to study group (administration of Liuwei dihuang pil s+celecoxib) and control group (administration of celecoxib alone). Preoperative, postoperative 6-and 12-month assessments of patients by the Lysholm Knee Scoring Scale, International Knee Documentation Committee (IKDC) Subjective Knee Form and MRI imaging were performed. RESULTS AND CONCLUSION:Postoperative Lysholm scores were significantly increased compared with preoperation in the study and control groups (P<0.05), and the scores of sections stair-climbing and instability and total scores of the Lysholm Knee Scoring Scale in the study group were superior to those in the control group (P<0.05). Postoperative IKDC scores were significantly increased compared with pre-operation in the study and control groups (P<0.05), and the scores of section flaccid leg and subjective function and total scores of the IKDC in the study group were superior to the control group (P<0.05). The enlargement rate of middle and proximal parts of grafts was significantly increased in the study group compared with the control group 6 months postoperatively (P<0.05), and the enlargement rate of middle and distal parts of grafts was significantly increased in the study group compared with the control group 12 months postoperatively (P<0.05). The tibial and femoral tunnel enlargement was observed in both groups at 6 and 12 months postoperatively (P<0.05). The enlargement rate of the tibial tunnel was significantly decreased in the study group compared with in the control group (P<0.05). The signal-noise ratio of the femoral end and middle part of grafts in the study group was significantly higher than that in the control group 6 months postoperatively (P<0.01). In contrast, the signal-noise ratio of the femoral end and middle part of grafts in the study group was significantly lower than that in the control group 12 months postoperatively (P<0.05). Our results indicate that kidney-tonifying herbs can improve the tendon-bone healing after ACLR.

Objective To explore the diagnosis and treatment of benign paroxysmal positional vertigo (BPPV) with bilateral positive Dix-Hallpike test.Methods This is a retrospective study based on the clinical data of BPPV patients diagnosed in the Dizziness Clinic of Changzheng Hospital from January 2012 to December 2012.Totally 490 patients with vertigo and nystagmus provoked by Dix-Hallpike maneuver were included in the present analysis.Results Among all the patients,55 (11.2％) of them presented with bilateral nystagmus by the provocative test.According to the type of nystagmus provoked by DixHallpike maneuver,the 55 patients can be divided into the following four categories.(1) Bilateral geotropic (n =16) and apogeotropic nystagmus (n =5):all these patients were diagnosed with horizontal canal BPPV and free of vertigo after head side-shaking exercise in supine position and Barbecue maneuver.(2) Bilateral predominant down-beating nystagmus (n =2):patients in this group were diagnosed with anterior canal BPPV,and got recovered after Kim maneuver.(3) Bilateral torsional up-beating geotropic nystagmus (n =20):after a lying-down test,6 of the patients manifested as vertical up-beating nystagmus and 14 patientsremained torsional up-beating nystagmus.The formerwere diagnosed with bilateral posterior canal BPPV,and were cured after bilateral PRM therapy,and the latter were diagnosed with horizontal canal BPPV,who were cured after Barbecue maneuver.(4) Torsional up-beating geotropic nystagmus on one side and down-beating nystagmus on the other side (n =12).The down-beating nystagrnus on the other side disappeared when the patients was firstly seated up with head down in 30 degrees for half an hour before second Dix-Hallpike maneuver.These patients were diagnosed with unilateral posterior canal BPPV and cured by PRM therapy.Conclusions It is common for vertigo patients with bilateral nystagmus induced by Dix-Hallpike test.The diagnoses should be made by the types of nystagmus provoked step by step before maneuver therapy.

OBJECTIVE: To analyze the feature of horizontal semicircular canal function at high and low-frequencies in Meniere's disease. METHOD: Thirty patients suffering from unilateral Meniere's disease were included in the research from 2013 June to 2014 June. Caloric test and video head impulse test were performed to evaluate the high low-frequency function of horizontal semicircular canal. RESULT: these patients were devided by the severity of unilateral weakness in caloric test. The gain value in video head impulse test, which reflects the high-frequency function of semicircular canal, were not different between the normal and mild abnormal group (P > 0.05), but were obviously different between the normal and mild-severe abnormal group, slight abnormal and mild-severe abnormal group (P < 0.05). Gain asymmetry, which reflects the high-frequency function of both side, has no difference between three groups (P > 0.05). CONCLUSION A part of Meniere's disease may have normal high, low-frequency function of horizontal semicircular canal. As patient suffering slight injury of low-frequency function, the high-frequency function keeps normal. As the injury of low-frequency function become mildly to severely, the damage of high-frequency function appears, but the symmetry still keeps balance.
Caloric Tests ; Head Impulse Test ; Humans ; Meniere Disease ; physiopathology ; Semicircular Canals ; physiopathology

Objective To study improvement of neural function by stereotaxic transplantation of adipose-derived stem cells (ADSC) into lateral cerebral ventricle after intracerebral hemorrhage in rats and its mechanism. Methods ADSC were cultured and proliferated in vitro, which had been marked with Brdu for 48 h before transplantation. The rat caudate nucleus hemorrhage (ICH) models were divided into 2 groups. ADSC were stereotaxically transplanted into the right lateral ventricles in ADSC group, and equal volume of saline was transplanted into control group. The score of neurological behavior were evaluated at modeling and 1, 3, 7, 14, 28 days after transplantation respectively.Double-staining immunofluorescence technique was used to detect Brdu-positive cells and the differentiation of neurons and astrocytes. In accordance with the instructions of TUNEL kit, cell apoptosis, and the expression of VEGF and angiogenesis were assayed. Results In vitro ADSC expressed undergo osteogenic and adipogenic differentiation. Compared with the control group, ADSC group had better motor function at 3, 7, and 14 days (P＜0. 05). Double-staining immunofluorescence showed mostly grafted Brdu-reactive ADSC had migrated to the hematoma zone, and some survivedand expressed Neun of Gfap. TUNEL analysis revealed that, 3 days after transplantation, the number of apoptotic cells in ADSC group was significantly less than in the control group (P＜0. 05). Three days after transplantation, VEGF expression levels in ADSC group were significantly higher than in the control group (P＜0. 05). Conclusion ADSC stereotaxially transplanted into the lateral ventricle can survive and differentiate into neuron-like cells. ADSC transplantation may reduce apoptosis and secret VEGF to promote the angiogenesis, and improve neural functional in intracerebral hemorrhage rats.

BACKGROUND:There are a lot of studies on isolation and culture methods of human placental chorionic-derived mesenchymal stem cells (hpcMSCs), but how to simply and efficiently harvest a large amount of primary MSCs has not been resolved. OBJECTIVE:To optimize the tissue explants method of isolating and culturing hpcMSCsin vitro. METHODS:Human placental chorionic villi were collected from full-term deliveries under aseptic condition and isolated by electric homogenizer. hpcMSCs were prepared by tissue explants method. The fluid and tissue of the primary culture flask and douching normal saline of the initial culture were centrifuged and prepared for secondary culture. RESULTS AND CONCLUSION: It saved time and effort to treat human placental chorionic villi with electric homogenizer, with good effects on tissue dispersion and removal of red blood cells. The average time of cell acquisition in initial culture and secondary culture was (17.73±1.14) and (10.03±1.30) days, respectively. The yields of primary cultured cells in initial culture and secondary culture were (6.97±0.98)×105 and (13.82±1.44)×105per Φ100 mm culture dish, respectively. The adherent cells showed fibroblast-cell-like shape, which were in parallel or circinate arrangement. Highly expressed CD73, CD105 and CD90 could be detected in the third generation of hpcMSCs, but CD34, CD45, CD14, CD19 and HLA-DR were negative. Following induction, alizarin red staining and oil red O staining produced a strong reaction in cells. In a word, the optimized method is a simple and efficient method for obtaining a large amount of primary hpcMSCs.

Objective To establish a cultivating method for obtaining a large number of P0 generation human placental chorionic-derived mesenchymal stem cells (hpcMSCs).Methods The hpcMSCs were isolated from human placental chorion.After primary culturing and culturing for seven days,the culture medium,the non-adherent tissue and the douching normal saline of the primary culture were centrifuged and re-cultured twice.Cell morphology was observed by an inverted microscope.CCK-8 was used to measure the cell growth curve.Flow cytometry was used to detect cell surface markers.Adipogenic and osteogenic differentiation kits were used to assess the cell differentiation potential.Results The obtained hpcMSCs were fibroblast-like adherent cells and (25.54±3.38)×106 cells were obtained per placenta.The total yield of the primary culture,secondary culture and tertiary culture were (11.73±2.09)×106,(11.12±1.42)×106 and (2.69±0.71)×106,respectively,and the incubation time were (12.00±0.64) d,(8.87±0.63) d and (12.33±0.80) d.There was significant differences in incubation time between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the incubation time of the tertiary culture had an increasing trend (P＞0.05).The yield per culture flask of the primary culture,secondary culture and tertiary culture were (1.12±0.15) × 106,(2.10±0.16)×106 and (1.04±0.16)×106,respectively.There was significant differences in the yield per culture flask between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the yield per culture flask of the tertiary culture had a decreasing trend (P＞0.05).There was no difference among the three cultures in the growth curve and the expression of surface markers,and the osteogenic and adipogenic differentiation were all positive.Conclusions The P0 generation hpcMSCs isolated from a choriocarcinoma sample can be doubled by the three cultures compared with the primary culture,which can provide plenty stem cell source for the regenerative medicine.

Objective To evaluate the effect of oxycodone on intestinal ischemia-reperfusion injury in rats and the role of autophagy.Methods Twenty-four healthy adult male Sprague-Dawley rats,aged 6-9 weeks,weighing 180-220 g,were divided into 4 groups (n =6 each) using a random number table method:sham group (group S),intestinal I/R group (group I/R),oxycodone group (group O) and oxycodone plus autophagy inhibitor 3-methyladenine (3-MA) group (group O+3-MA).Intestinal I/R was induced via clamping the superior mesenteric artery for 1 h,followed by 2-h reperfusion in all the groups except for group S.Oxycodone 0.5 mg/kg was injected via caudal vein at 15 min before ischemia in group O and group O+3-MA.3-MA 30 mg/kg was intraperitoneally injected at 10 min before ischemia in group O+3-MA.Rats were gavaged with fluorescein-isothiocyanate-conjugated dextran (FITC-dextran) immediately before ischemia.Blood samples were collected from the cardiac apex at 2 h of reperfusion to detect the level of serum FITC-dextran.Blood samples were collected from the cardiac apex at 2 h of reperfusion to measure the concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) in serum by enzyme-linked immunosorbent assay.Small intestinal tissues were obtained at 2 h of reperfusion for examination of the pathological changes and for determination of the expression of occludin,Beclin-1 and microtubule-associated protein 1 light chain 3 (LC3) (by Western blot).Intestinal damage was assessed and scored according to Chiu.The ratio of LC3 Ⅱ expression to LC3 Ⅰ expression (LC3 Ⅱ / Ⅰ) was calculated.Results Compared with group S,the Chiu's score,levels of serum FITC-dextran,TNF-α and IL-1β and LC3 Ⅱ/Ⅰ ratio were significantly increased,the expression of Beclin-1 was up-regulated,and the expression of occludin was down-regulated in group I/R (P＜0.05).Compared with group I/R,the Chiu's score and levels of serum FITC-dextran,TNF-α and IL-1β were significantly decreased,LC3 Ⅱ / Ⅰ ratio was increased,and the expression of occludin and Beclin-1 was up-regulated in group O (P＜0.05),and no significant change was found in the parameters mentioned above in group O+ 3-MA (P＞0.05).Compared with group O,the Chiu's score and levels of serum FITC-dextran,TNF-α and IL-1β were significantly increased,LC3 Ⅱ / Ⅰ ratio was decreased,and the expression of occludin and Beclin-1 was down-regulated in group O + 3-MA (P＜ 0.05).Conclusion Oxycodone can ameliorate intestinal I/R injury,and the mechanism may be related to enhancing autophagy in intestinal tissues of rats.

Adipose tissue is a readily available source of adult stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications.Peptide hydrogel is a novel biomaterial which provides three-dimensional microenvironments for a variety of cells for tissue grafting.In this study,adipose-derived stem cells (ADSCs) were isolated from rats,seeded into the peptide hydrogel polymer scaffolds and cultured in Neurobasal (NB) media supplemented with B27,basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF).Ten days after the culture,some cells were expanded into clonal populations in which the expression of both Nestin and Brdu was detected but only Brdu expression was detected in the cells that were not expanded into clonal populations.Our results suggested that ADSCs in peptide hydrogel polymer scaffolds can be induced to differentiate into cells capable of expressing the neuron-associated markers,self-renewal and self-propagation.

Background: and Purpose Non-high-density lipoprotein cholesterol (non-HDL-C), which represents the total cholesterol content of all pro-atherogenic lipoproteins, has recently been included as a new target for lipid-lowering therapy in high-risk atherosclerotic patients in multiple guidelines. Herein, we aimed to explore the relationship between non-HDL-C level and the efficacy and safety of ticagrelor-aspirin versus clopidogrel-aspirin in preventing stroke recurrence. Methods: This study comprised a post hoc analysis of the CHANCE-2 (Ticagrelor or Clopidogrel in High-Risk Patients with Acute Nondisabling Cerebrovascular Events II) trial, from which 5,901 patients with complete data on non-HDL-C were included and categorized by median non-HDL-C levels, using a cutoff of 3.5 mmol/L. The primary efficacy and safety outcomes were recurrent stroke and severe or moderate bleeding within 90 days. Results: Ticagrelor-aspirin significantly reduced the risk of recurrent stroke in patients with low non-HDL-C (71 [4.8%] vs. 119 [7.7%]; adjusted hazard ratio [HR] 0.54; 95% confidence interval [CI], 0.40–0.74), but not in those with high non-HDL-C (107 [7.3%] vs. 108 [7.6%]; adjusted HR, 0.88; 95% CI, 0.67–1.16), compared with clopidogrel-aspirin (P for interaction=0.010). When analyzed as a continuous variable, the benefit of ticagrelor-aspirin for recurrent stroke decreased as non-HDL-C levels increased. No significant differences in the treatment assignments across the non-HDL-C groups were observed in terms of the rate of severe or moderate bleeding (5 [0.3%] vs. 8 [0.5%] in the low non-HDL-C group; 4 [0.3%] vs. 2 [0.1%] in the high non-HDL-C group; P for interaction=0.425). Conclusion CHANCE-2 participants with low non-HDL-C levels received more clinical benefit from ticagrelor-aspirin versus clopidogrel-aspirin compared to those with high non-HDL-C, following minor ischemic stroke or transient ischemic attack.