bHLH transcription factors Hes1, Ascl1 and Oligo2 exhibit different expression patterns in the process of physiological electric fields-induced neuronal differentiation.

BACKGROUND: Recent studies have shown that the expression of bHLH transcription factors Hes1, Ascl1, and Oligo2 has an oscillating balance in neural stem cells (NSCs) to maintain their self-proliferation and multi-directional differentiation potential. This balance can be disrupted by exogenous stimulation. Our previous work has identified that electrical stimulation could induce neuronal differentiation of mouse NSCs. METHODS: To further evaluate if physiological electric fields (EFs)-induced neuronal differentiation is related to the expression patterns of bHLH transcription factors Hes1, Ascl1, and Oligo2, mouse embryonic brain NSCs were used to investigate the expression changes of Ascl1, Hes1 and Oligo2 in mRNA and protein levels during EF-induced neuronal differentiation. RESULTS: Our results showed that NSCs expressed high level of Hes1, while expression of Ascl1 and Oligo2 stayed at very low levels. When NSCs exited proliferation, the expression of Hes1 in differentiated cells began to decrease and oscillated at the low expression level. Oligo2 showed irregular changes in low expression level. EF-stimulation significantly increased the expression of Ascl1 at mRNA and protein levels accompanied by an increased percentage of neuronal differentiation. What's more, over-expression of Hes1 inhibited the neuronal differentiation induced by EFs. CONCLUSION: EF-stimulation directed neuronal differentiation of NSCs by promoting the continuous accumulation of Ascl1 expression and decreasing the expression of Hes1.

Clinical Features of Non-Alcoholic Fatty Liver Disease in the Non-Lean Population.

INTRODUCTION: The prevalence of non-alcoholic fatty liver disease (NAFLD) in non-lean patients is significantly increased, and obesity significantly increases the risk of cirrhosis and HCC in NAFLD patients. However, whether there is a difference in clinical manifestations of NAFLD between overweight and obesity remains unclear. The objective of this study was to assess the clinical and histological features of NAFLD among a non-lean population. METHODS: Current study enrolled consecutive non-lean (body mass index [BMI] >23 kg/m2) patients with NAFLD and available liver biopsy results. Patients were stratified by BMI into two groups for the comparison of their clinical and histological variables, which included the overweight (BMI 23∼<28 kg/m2) and the obese (BMI ≥28 kg/m2). Risk factors for moderate to severe fibrosis (stage >1) were also analyzed through the logistic regression model. RESULTS: Among 184 non-lean patients with metabolic-associated fatty liver disease enrolled, 65 and 119 were overweight and obese, respectively. Patients in the obesity group had a significantly lower level of gamma-glutamyl transpeptidase, higher levels of platelet, glucose, prothrombin time, and more common of moderate to severe inflammatory activity when compared to those in the overweight group. However, a significant low frequency of moderate to severe fibrosis was found in the obesity group versus the overweight group (19.33% vs. 40.00%, p = 0.002). Binary logistics regression analysis of fibrosis found that aspartate transaminase (AST), BMI, alanine transaminase (ALT), and cholesterol (CHOL) were independent predictors for moderate to severe fibrosis in non-lean patients with NAFLD. Compared with the traditional fibrosis-4 (AUC = 0.77) and aminotransferase to platelet ratio index (AUC = 0.79) indexes, the combined index based on AST, BMI, ALT, and CHOL was more accurate in predicting moderate to severe fibrosis in non-lean patients with NAFLD (AUC = 0.87). CONCLUSIONS: Clinical and histological features differed between obesity and overweight patients with NAFLD. When compared to the traditional serum markers, the combination index including AST, BMI, ALT, and CHOL provided a better model to predict moderate to severe fibrosis in non-lean patients with NAFLD.

Totally laparoscopic total gastrectomy using the "enjoyable space" approach coupled with self-pulling and latter transection reconstruction versus laparoscopic-assisted total gastrectomy for upper gastric cancer: short-term outcomes.

Introduction: With the development of minimally invasive surgery in recent years, totally laparoscopic total gastrectomy (TLTG) has attracted more attention. Aim: To introduce the more comprehensive "enjoyable space" approach coupled with the self-pulling and latter transaction (SPLT) reconstruction technique to perform TLTG and investigate its safety and feasibility. Material and methods: Ninety-seven patients with primary upper gastric cancer underwent laparoscopic radical total gastrectomy between January 2020 and December 2020. Among these patients, 46 underwent laparoscopic-assisted total gastrectomy (LATG), and 51 underwent TLTG. We compared the clinicopathological characteristics, surgical outcomes and postoperative complications between the two groups. Results: There were no significant differences in the clinicopathological characteristics between the two groups (p > 0.05). However, the TLTG group had a slightly lower mean operative time and mean blood loss than the LATG group (p < 0.05 each). Although there were similarities in the mean times to first flatus, liquid diet, and soft diet, the duration of hospital stay was significantly reduced in the TLTG group (p < 0.05). No significant differences in overall complications and E-J-related complications were found between the two groups (15.2% vs. 25.4%, p > 0.05). Conclusions: TLTG is a safe and feasible procedure for treating upper gastric cancer. The enjoyable space approach in conjunction with SPLT reconstruction is an appropriate comprehensive technique with several advantages over LATG.

Arylpentazoles with surprisingly high kinetic stability.

The more-than-one-century-old arylpentazoles can only be used in situ in generating the pentazole anion due to their unfavourable kinetic stability. We successfully increased the N2-leaving barrier to reach hitherto the highest value of 40.83 kcal mol-1 at the CBS-QB3 level via a newly proposed co-stabilization method, making the broader applications of arylpentazoles feasible.

Combination of electrical stimulation and bFGF synergistically promote neuronal differentiation of neural stem cells and neurite extension to construct 3D engineered neural tissue.

OBJECTIVE: The construction of in vitro three-dimensional (3D) neural tissue has to overcome two main types of challenges: (1) How to obtain enough number of functional neurons from stem cells in 3D culture; (2) How to wire those lately developed neurons into functional neural networks. Here, we describe the potential of using direct current (DC) electric field (EF) together with basic fibroblast growth factor (bFGF) synergistically in promoting neural stem cell (NSC) neuronal differentiation following by directing neurite outgrowth in the 3D neural tissue construction. APPROACH: By adjusting the electrical stimulation setup in this study, long-term electrical stimulation could be present in vitro. At an EF strength of 150 mV mm-1, cell responses, including cell viability, neuronal differentiation, cell morphology, the length of neuronal processes, synaptic structure and neural network formation, were quantified and analyzed. MAIN RESULTS: Analysis revealed that NSCs showed no significant cell death after certain EF treatments. EF-stimulated NSCs in 3D Matrigel mainly differentiated into neurons, but unlike NSCs in two-dimensional conditions, their processes were flat and stunted. When combined with bFGF, EF stimulation provided appropriate bioactive cues to establish engineered neural tissue with a proper neuronal cell number, highly branched neurites, and a well-developed neuronal network. SIGNIFICANCE: It is for the first time the synergistic effects of EF and bFGF stimulation have been evaluated in inducing the differentiation of NSCs into neurons and the acquisition of long neurites in a culture environment of in vitro 3D model. These optimized conditions may allow a well-developed neuronal network to be established within hydrogel droplets.

Laparoscopic D2 plus complete mesogastrium excision using the "enjoyable space" approach versus conventional D2 total gastrectomy for local advanced gastric cancer: short-term outcomes.

INTRODUCTION: Laparoscopy-assisted radical total gastrectomy is technically demanding. AIM: To introduce the "enjoyable space" approach to achieve D2 plus complete mesogastrium excision (CME) and to investigate its safety and feasibility. MATERIAL AND METHODS: Between January 2015 and December 2017, 165 patients with primary advanced upper gastric cancer underwent laparoscopy-assisted radical total gastrectomy. Among these patients, 81 underwent conventional D2 total gastrectomy (D2 group) and 84 underwent D2 plus CME total gastrectomy (D2 + CME group). Clinicopathological characteristics, surgical outcomes and postoperative complications were compared between the two groups. RESULTS: There were no significant differences between the two groups (p > 0.05) in clinicopathological characteristics. However, the D2 + CME group had a longer mean operative time, lower mean blood loss and slightly higher mean number of retrieved lymph nodes (LNs) than the D2 group (p < 0.05 each). The mean time to first flatus, liquid diet, and soft diet and the duration of hospital stay were similar between the two groups (p > 0.05 each). No significant difference in postoperative complication rates was found between the groups (16.0% vs. 9.5%, p > 0.05). CONCLUSIONS: The "enjoyable space" approach is an option to achieve D2 + CME, and its safety and feasibility over conventional method are confirmed with lower intraoperative blood loss and more harvested LNs.

Laparoscopic perigastric mesogastrium excision technique for radical total gastrectomy.

INTRODUCTION: Complete mesogastrium excision (CME) of the perigastric mesogastrium and dissection of lymph nodes (LNs) no. 10 and no. 11 remain technically challenging aspects of laparoscopic radical total gastrectomy (LRTG) plus CME. To address some of these difficulties, we introduced the laparoscopic perigastric mesogastrium excision technique (LPMET) and the concept of the "enjoyable space" to partly modify the procedures of conventional radical surgery and characterize the perigastric space and the surgical plane as well as its boundaries. AIM: To introduce the laparoscopic perigastric mesogastrium excision technique (LPMET) and the "enjoyable space" when undergoing laparoscopic radical total gastrectomy. MATERIAL AND METHODS: From July 2016 to June 2017, 79 cases of upper gastric cancer that were treated by laparoscopic D2 gastrectomy plus CME were investigated. The retrospective database included the patient characteristics, intraoperative and postoperative outcomes, and morbidity and mortality rates depending on the completeness of their medical records. RESULTS: Laparoscopic D2 gastrectomy plus CME was successfully performed in all 79 cases. The mean surgical time was 232.5 ±46.0 min, and the intraoperative blood loss was 67.6 ±52.3 ml. A total of 2245 LNs were retrieved (mean 28.1 ±10.8 retrieved from each specimen). The mean postoperative hospital stay was 10.3 ±1.6 days. The postoperative morbidity rate was 17.7%. After a median follow-up period of 12 months, one patient experienced liver metastasis; of the other 78 patients, none died or experienced tumor recurrence or metastasis. CONCLUSIONS: Laparoscopic perigastric mesogastrium excision technique and the "enjoyable space" could be a novel, minimally invasive approach and space to achieve CME and provide benefit for the dissection of LNs no. 10 and no. 11.

Ascl1 Regulates Electric Field-Induced Neuronal Differentiation Through PI3K/Akt Pathway.

Directing differentiation of neural stem/progenitor cells (NSCs/NPCs) to produce functional neurons is one of the greatest challenges in regenerative medicine. Our previous paper has confirmed that electrical stimulation has a high efficiency of triggering neuronal differentiation by using isolated filum terminale (FT)-derived NPCs. To further clarify the intrinsic molecular mechanisms, protein-protein interaction (PPI) network analysis was applied to pinpoints novel hubs in electric field (EF)-induced neuronal differentiation. In this study, siRNA transfection of Achaete-scute homolog 1 (Ascl1) in NPCs or NPCs was followed by direct current stimulation at 150 mV/mm. Neuronal differentiation rate and protein expression level were analyzed after 7 or 14 days of electrical stimulation. The data showed that the expression level of Ascl1 was enhanced by electrical stimulation and positively correlated to EF strength. Moreover, we identified that the expression of Ascl1 positively regulated neuronal differentiation of NPCs and can be up-regulated by EF-stimulation through the activation of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathway. Therefore, this study provides new insights into the role of Ascl1 and its relevant PI3K/Akt pathway in regulating of EF-induced neuronal differentiation and pointed out that continuous expression of Ascl1 in NPCs is required for EF-induced neuronal differentiation.

Electric field stimulation induced neuronal differentiation of filum terminale derived neural progenitor cells.

Adult filum terminale (FT) is an atypical region from where multipotent neural progenitor cells (NPCs) have been isolated. However, poor neuronal differentiation rate of FT-NPCs currently limits their clinical applications. Using custom-designed electric fields (EFs), this study sets up a method to significantly improve neuronal differentiation rate of rat FT-NPCs in vitro. We investigated the influence of EF strength on rat FT-NPCs differentiation. By adding reasonable strength of EF to FT-NPCs, our data shows a significant increase in neuronal differentiation rate. The present innovation provides a novel method of directional differentiation and efficient production of neurons from FT-NPCs in vitro. This improved approach for inducing neuronal differentiation can be applied to future research on autoplastic transplantation.

Remediation of soil co-contaminated with petroleum and heavy metals by the integration of electrokinetics and biostimulation.

Successful remediation of soil co-contaminated with high levels of organics and heavy metals is a challenging task, because that metal pollutants in soil can partially or completely suppress normal heterotrophic microbial activity and thus hamper biodegradation of organics. In this study, the benefits of integrating electrokinetic (EK) remediation with biodegradation for decontaminating soil co-contaminated with crude oil and Pb were evaluated in laboratory-scale experiments lasting for 30 days. The treated soil contained 12,500 mg/kg of total petroleum hydrocarbons (TPH) and 450 mg/kg Pb. The amendments of EDTA and Tween 80, together with a regular refreshing of electrolyte showed the best performance to remediate this contaminated soil. An important function of EDTA-enhanced EK treatment was to eliminate heavy metal toxicity from the soil, thus activating microbial degradation of oil. Although Tween 80 reduced current, it could serve as a second substrate for enhancing microbial growth and biodegradation. It was found that oil biodegradation degree and microbial numbers increased toward the anode and cathode. Microbial metabolism was found to be beneficial to metal release from the soil matrix. Under the optimum conditions, the soil Pb and TPH removal percentages after 30 days of running reached 81.7% and 88.3%, respectively. After treatment, both the residual soil Pb and TPH concentrations met the requirement of the Chinese soil environmental quality standards.

Pretreatment of old-age landfill leachate by microwave-assisted catalytic oxidation in the presence of activated carbon.

Landfill leachate is posing an ever-greater environmental hazard. Recently, a process for purification combining activated carbon, microwave (MW) and Fenton oxidation has drawn much attention. In this study, the effectiveness of this process for the pretreatment of an old-age landfill leachate was tested. The effects of various parameters were investigated and the optimal condition included as follows: MW energy density, 6 W/mL; MW power, 300 W; radiation time, 8 min; H2O2 dosage, 0.1 mol/L; Fe(2+)-EDTA dosage, 0.02 mol/L; granular activated carbon (GAC) dosage, 6 g/L. Within the present experimental condition applied, the chemical oxygen demand (COD) removal reached 56.5%, and the ratio of 5-day biochemical oxygen demand to chemical oxygen demand (BOD5/COD) was enhanced from 0.122 to 0.462. Comparing with GAC, MW and Fenton alone or the combinations of any two of them, MW/Fenton/GAC displayed superior treatment efficiency. The MW/Fenton/GAC process is believed to be a promising pretreatment technology for biorefractory old-age landfill leachate.

Treatment of cholecystitis with Chinese herbal medicines: a systematic review of the literature.

AIM: To analyze the literature on the use of Chinese herbal medicines for the treatment of cholecystitis. METHODS: The literature on treatment of cholecystitis with traditional Chinese medicines (TCM) was analyzed based on the principles and methods described by evidence-based medicine (EBM). Eight databases including MEDLINE, EMbase, Cochrane Central (CCTR), four Chinese databases (China Biological Medicine Database, Chinese National Knowledge Infrastructure Database, Database of Chinese Science and Technology Periodicals, Database of Chinese Ministry of Science and Technology) and Chinese Clinical Registry Center, were searched. Full text articles or abstracts concerning TCM treatment of cholecystitis were selected, categorized according to study design, the strength of evidence, the first author's hospital type, and analyzed statistically. RESULTS: A search of the literature published from 1977 through 2009 yielded 1468 articles in Chinese and 9 in other languages; and 93.92% of the articles focused on clinical studies. No article was of level I evidence, and 9.26% were of level II evidence. The literature cited by Science Citation Index (SCI), MEDLINE and core Chinese medical journals accounted for 0.41%, 0.68% and 7.29%, respectively. Typically, the articles featured in case reports of illness, examined from the perspective of EBM, were weak in both quality and evidence level, which inconsistently conflicted with the fact that most of the papers were by authors from Level-3 hospitals, the highest possible level evaluated based on their comprehensive quality and academic authenticity in China. CONCLUSION: The published literature on TCM treatment of cholecystitis is of low quality and based on low evidence, and cognitive medicine may functions as a useful supplementary framework for the evaluation.

Diagnostic value of sentinel lymph node biopsy in gastric cancer: a meta-analysis.

BACKGROUND: The possible application and validity of the sentinel lymph node (SLN) concept in gastric cancer (GC) is still debated. A systematic review to evaluate the diagnostic value of SLN biopsy (SLNB) in GC is urgently needed. METHODS: A systematic review of relevant literatures was performed in PubMed, Embase, and The Cochrane Library. A random-effect model was used to pool the data, and subgroup analysis was used to explain the heterogeneities. RESULTS: A total of 38 included studies (2,128 patients) were included. The pooled SLN identification rate, sensitivity, negative predictive value, and accuracy were 93.7% (95% confidence interval [95% CI]: 91.1-95.6%), 76.9% (95% CI: 71.6-81.4%), 90.3% (95% CI: 86.9-92.9%), and 92.0% (95% CI: 89.9-93.7%), respectively. Subgroup analysis showed that early T stage, combined tracers, submucosal injection method, conventional open surgery, and usage of immunohistochemistry were associated with higher SLN identification rate and sensitivity. CONCLUSIONS: SLNB in GC is technically feasible with an acceptable sensitivity. However, further studies are needed to confirm the best procedure and standard criteria.

High efficient isolation and systematic identification of human adipose-derived mesenchymal stem cells.

BACKGROUND: Developing efficient methods to isolate and identify human adipose-derived mesenchymal stem cells (hADSCs) remains to be one of the major challenges in tissue engineering. METHODS: We demonstrate here a method by isolating hADSCs from abdominal subcutaneous adipose tissue harvested during caesarian section. The hADSCs were isolated from human adipose tissue by collagenase digestion and adherence to flasks. RESULTS: The yield reached around 1 × 10(6) hADSCs per gram adipose tissue. The following comprehensive identification and characterization illustrated pronounced features of mesenchymal stem cells (MSCs). The fibroblast-like hADSCs exhibited typical ultrastructure details for vigorous cell activities. Karyotype mapping showed normal human chromosome. With unique immunophenotypes they were positive for CD29, CD44, CD73, CD105 and CD166, but negative for CD31, CD34, CD45 and HLA-DR. The growth curve and cell cycle analysis revealed high capability for self-renewal and proliferation. Moreover, these cells could be functionally induced into adipocytes, osteoblasts, and endothelial cells in the presence of appropriate conditioned media. CONCLUSION: The data presented here suggest that we have developed high efficient isolation and cultivation methods with a systematic strategy for identification and characterization of hADSCs. These techniques will be able to provide safe and stable seeding cells for research and clinical application.

[Integrated traditional Chinese and Western medicine versus Western medicine in treatment of arteriosclerosis obliterans: a systematic review of randomized controlled trials].

BACKGROUND: The conventional therapy for arteriosclerosis obliterans (ASO) is Western medicine. However, it has some adverse effects and does not respond to some patients, and it is also very expensive. OBJECTIVE: To evaluate the efficacy of integrated traditional Chinese (TCM) and Western medicine (WM) in treatment of ASO. SEARCH STRATEGY: Electronic and manual searches were conducted and the searches ended on May 20, 2009. INCLUSION CRITERIA: We included randomized controlled trials (RCT) evaluating integrated TCM and WM (as treatment group) versus WM used alone (as control group), and no language limits were set. DATA EXTRACTION AND ANALYSIS: Selection of trials for inclusion, assessment for methodological quality, data extraction and data syntheses were conducted according to protocol of a Cochrane systematic review by the authors. RESULTS: Thirteen RCT were included, which encompassed a total of 968 patients. The results showed that all of the 13 included trials did not report mortality rate of ASO. The studies displayed that the amputation rate in the treatment group was lower than that in the control group, but there was no statistical significance. Ten studies adopted inefficiency analysis and 2 of them showed that the ineffective rate in the treatment group was lower than that in the control group, and the relative risk (RR) and 95% CI were 0.36 [0.13, 0.99]. We performed descriptive analysis on other 8 studies; analyses of secondary outcomes such as intermittent claudication, ankle brachial index, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) showed that integrated TCM and WM therapy was more effective than WM treatment alone; one study showed that WM was better than integrated TCM and WM therapy in decreasing the content of fibrinogen. All of the included trials did not report any critical adverse reactions occurred in the treatment group. CONCLUSION: The current evidence shows that integrated TCM and WM therapy is safe and effective in treating ASO, and tends to reduce amputation rate, improve intermittent claudication, decrease the levels of fibrinogen, HDL and LDL, and increase ankle brachial index, without obvious adverse reactions. Due to the low methodological quality of trials included, more prospective, multicenter, large-scale, high-quality RCTs are needed.

[Experimental study on spinal fusion induced by hBMP-4 gene modified tissue engineered bone].

OBJECTIVE: To evaluate the efficacy of hBMP-4 gene modified tissue engineered bone graft in the enhancement of rabbit spinal fusion and find an ideal kind of substitute for the autograft bone. METHODS: Rabbit BMSCs were cultured and transfected with AAV-hBMP-4 using different MOI value. The optimal MOI value were determined by observing cell's morphology change. BMSCs were then transfected with AAV-hBMP4 and AAV-EGFP respectively, following which the transfected cells were evenly suspended in a collagen sponge I, and implanted to either side of the L5,6 intertransverse spaces posterolateral in the New Zealand rabbits to induce spinal fusion. Fourteen rabbits were randomly divided into 2 groups. Group 1: AAV-hBMP-4 transfected BMSCs in the right side (hBMP-4 side) and autograft bone in the left side. Group 2: AAV-hBMP-4 transfected BMSCs in the right side (hBMP-4 side) and AAV-EGFP transfected BMSCs in the left side (EGFP side). Radiographs and three-dimensional CT of the spine, manual palpation, gross and histological examination of the fusion masses for all the animals were performed subsequent to animals having been sacrificed at 12 weeks after surgery. RESULTS: Evaluation has been taken in 12 New Zealand rabbits delivered into 2 groups which meet the criterion after operation. Eleven in 12 implemented sides involved hBMP-4 achieved bony fusion, to which 5 in 6 autografted sides was similar. But only 2 in 6 sides in EGFP-group achieved bony fusion meanwhile. Three-dimensional CT scan and palpation also evidenced the results. Bone formation was observed obviously on specimen both in hBMP4 sides and autografted ones. EGFP-group also got bony integration, but the quantity was small. CONCLUSION: Tissue-engineered bone graft constructed from application of hBMP4 is a fine substitute for autograft. Effective enhancement of bony integration in spinal fusion surgery has been evidenced in vivo.

Co-transplantation of bFGF-expressing amniotic epithelial cells and neural stem cells promotes functional recovery in spinal cord-injured rats.

We have previously demonstrated that amniotic epithelial cells (AECs) can enhance survival and neural differentiation of neural stem cells (NSCs) when co-cultured in basal media. In addition, the presence of basic fibroblast growth factor (bFGF) enhances this AEC function. The aim of the present study was to extend those findings and investigate whether AECs modified with the bFGF gene will also enhance NSCs survival and neural differentiation in vivo and promote repair of the injured spinal cord. Female Wistar rats were used for a contusive spinal cord injury (SCI) model. Contusive SCIs were induced using a weight-drop device at levels T9-T11. Seven days following contusion, rats received grafts of NSCs only, NSCs with AECs/pLEGFP-hbFGF, or NSCs with AECs/pLEGFP-C1 into the injured region. Significant locomotor improvement was observed in the NSCs/AECs co-graft group beginning at 3 weeks compared with the NSCs or NaCl only groups. These results were confirmed and extended in an electrophysiological analysis. An immunohistological analysis revealed that AECs/pLEGFP-hbFGF promoted the survival (vs NaCl group: 194+/-9.17 vs 103.6+/-13.05) and neural differentiation (vs NaCl group: 14.24+/-1.11 vs 7+/-0.63) of co-transplanted NSCs. We also confirmed that AECs could promote the survival of host neurons. These results suggest that AECs/pLEGFP-hbFGF improve the NSCs survival and differentiation microenvironment and may be useful as a source of sustained trophic supported to improve NSCs differentiation into neurons in vivo. These findings suggest that a cograft of AECs/pLEGFP-hbFGF and NSCs may have benefits for SCI.

[One-stage single balloon multiple expansions percutaneous kyphoplasty: report of 18 cases].

OBJECTIVE: To observe the effect and safety of one-stage single balloon multiple expansions percutaneous kyphoplasty (PKP) in treatment of osteoporotic vertebral compressive fracture (OVCF) and spinal tumor. METHODS: One-stage single balloon multiple expansions PKP was performed on 18 patients, 5 males and 13 females, aged 68.77 (44 - 80), with 45 vertebrae, including 11 case of OVCF (with 29 diseased vertebrae), 11 cases of multiple vertebral fracture (with 19 diseased vertebrae), 5 cases of multiple myeloma (with 12 diseased vertebrae), and 2 case of spinal metastatic tumor (with 4 diseased vertebrae), the vital signs were record during operation. Pain relief and functional recovery were evaluated with visual analogue pain scale (VAS) and Oswestry disability index (ODI) scaling, and restoration of vertebral height and Cobb angle were evaluated by X-ray examination. Follow-up was conducted by telephone survey or clinic consults for 12.3 months (6 - 18 months). RESULTS: Operation was successfully performed on all patients with an operative time of 29.3 min (55 - 127 min) per vertebra. The average pressure of the balloon expansion was 165 Psi (87 - 210 Psi), and the average balloon expanded volume was 3.25 ml (1.5 - 4 ml). A balloon was expanded 2 approximately 5 times in one operation with the average of 2.94 expanded times. The bone cement volume injected was 3.95 ml (2 - 8.5 ml) per vertebra. Epidural cement leakage was seen in 1 vertebra in 1 case and paraspinal leakage was seen in two vertebrae in 1 case. The VAS and ODI scoring of these patients were both decreased significantly after operation. Both the anterior height and midline height of vertebrae were significantly improved. The pain relief and functional recovery were substantial and maintained to the last follow-up without any re-collapse or adjacent level fracture. CONCLUSION: one-stage single balloon multiple expansions PKP is effective, economic and safe in treatment of multi-level OVCF and spinal tumor.

Preliminary clinical outcomes of percutaneous kyphoplasty with Sky-bone expander.

BACKGROUND: Percutaneous kyphoplasty (PKP) using balloon expander has been proved to be effective in the treatment of painful vertebral compression fractures. Recently, Sky-bone expander, an alternative bone expander for PKP has been developed. The purpose of this study was to show our preliminary clinical outcomes of PKP with Sky-bone expander. METHODS: PKP with Sky-bone expander was performed in 25 patients (30 vertebrae). The operation time, bleeding volume, cement volume injected were recorded. The pain and functional activities of the patients before and after the operation were compared using Wilcoxon signed-rank test. The cement distribution in the vertebrae, vertebral height restoration, and kyphosis correction after the procedure were evaluated by radiography. The pre- and post-operative absolute values of the vertebral height and kyphotic angle were compared by paired-sample t test. All the patients were followed up by telephone or clinic consulting after being discharged from our hospital. RESULTS: The procedure was performed successfully in all the patients. Bipedicular injection was used in 2 of the patients, and unipedicular injection was made in the others. The operation time ranged from 25 to 120 minutes (45 minutes per vertebra on average). The average bleeding volume was about 20 ml. Polymethylmethacrylate 1.5-5.0 ml (mean, (3.15+/-0.78) ml) was injected through each pedicle into all the patients except one, who received calcium sulphate 3.5 ml instead. The patients were followed up for 12-15 months (13.5 months on average). The mean visual analogue scale (VAS) score, Oswestry Disability Index, anterior, midline, and posterior vertebral height, and kyphotic angle of the patients were improved significantly at the end of the follow-up compared with those before the operation. (2.5+/-1.3, 35.1%, (20.94+/-6.15) mm, (20.26+/-4.59) mm, (26.72+/-3.49) mm, and 8.2 degrees vs. 8.5+/-1.9, 61.2%, (19.11+/-6.72) mm, (15.88+/-5.73) mm, (25.78+/-3.67) mm, and 17.3 degrees; all P<0.05). The cement distribution with unipedicular injection was mostly limited within the injection site in the vertebral body. Cement extravasation was seen at ten levels (33.3%). CONCLUSIONS: PKP with Sky-bone expander is an effective and relatively safe alternative to the PKP using balloon expander. It can relieve pain, improve physical function, and restore the height of the collapsed vertebrae, but the cement extravasation is unsolved.

Enhanced neural differentiation of neural stem cells and neurite growth by amniotic epithelial cell co-culture.

Amniotic epithelial cells (AECs) were reported to show a neuroprotective effect on neurons, but there was no direct evidence for a functional relationship between neural stem cells (NSCs) and AECs. The aim of this study was to determine whether AECs could stimulate differentiation and expand neurogenesis of NSCs, and whether the roles were due to a diffusible factor or required direct cell-cell contact. AECs were isolated from rat amnion on E14-16 and NSCs were isolated from neocortical tissue. The growth and differentiation of NSCs were compared under different conditions. The results showed that NSCs cultured with FGF-2 proliferated and formed floating neurospheres while those grown in B27 without FGF-2 failed to thrive. Those grown either with AEC conditioned medium or in transwells showed significantly improved survival. Moreover, the neural differentiation and length of neurite were greater in exogenous FGF groups when NSCs were allowed direct contact with AECs. Western blotting, immunocytochemistry and RT-PCR indicated that rat AECs could secrete NT-3 and BDNF. Furthermore, the presence of FGF-2 enhanced the function of AECs. These findings identified that AECs may be regarded as a critical component of NSCs niche and suggested that direct cell-to-cell contact may provide additional and independent support. Such information would circumvent the need for AECs-NSCs co-culture and could potentially facilitate the production of neurons for future clinical applications.