An Optimized Method for Adipose Stromal Vascular Fraction Isolation and its Application in Fat Grafting.

BACKGROUND: The stromal vascular fraction (SVF) derived from adipose tissue contains heterogeneous cell populations and has enormous potential for clinical therapy. There are two main methods for SVF isolation: enzymatic isolation and mechanical isolation, both of which have shortcomings. In this study, optimized conditions for the isolation of high-quality SVF were established, and applications in fat grafting were evaluated. METHODS: Adipose tissue was chopped into small pieces and then ground into an erosive shape using a syringe. The pieces were digested with 0.15% type II collagenase for 35 min at 37 °C. After centrifugation, the pellets were resuspended in DMEM and passed through a 100-µm strainer. The filtered cells were analyzed by flow cytometry. The fat graft was enriched with isolated SVF and subcutaneously transplanted into nude mice. Three weeks after transplantation, grafts were isolated, and H&E staining, immunocytochemistry, and western blotting were conducted. RESULTS: The harvested SVF cells reached > 2 × 106/ml of adipose tissue within 90 min of operation. The number of CD34+ ADSCs in our SVF pellets was > 6 × 105/ml of adipose tissue, which has the potential for differentiating into osteoblasts, adipocytes, and chondrocytes. Freshly collected adipose tissue is better for SVF isolation, and isolated SVF should also be kept at 4 °C and used as soon as possible. SVF may promote revascularization after fat grafting. The adipose tissue of an SVF co-transplanted group had an integral structure, clear capillaries, and higher VEGF expression. SVF co-transplantation inhibited adipose cell apoptosis. CONCLUSION: Our study provides an efficient procedure for SVF isolation, its application in fat grafting, and possible underlying mechanisms. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Bioresorbable Vascular Scaffold: A Focused Review on Development and Preclinical Studies / 中国医疗器械杂志

Bioresorbable vascular scaffolds(BVS) are new treatment strategies of percutaneous coronary intervention. They have been introduced to overcome limitations of bare metal stents (BMS) and drug-eluting stents(DES), since they provide temporary scaffolding and then disappear, liberate the treated vessel from cage. In this article, we review the current status and problems of BVS, various tests required before gaining regulatory approval for clinical use.

Comparison of organ coefficients and expression of hypoxia-related genes in the Bama and Juema minipigs / 中国比较医学杂志

Objective To compare the organ coefficients and expressions of hypoxia-related genes in Bama and Juema pigs.Method Real-time quantitative PCR was used to detect the changes of hypoxia gene expressions in the heart,liver,spleen,lung,and kidney of Juema and Bama miniature pigs.Results The organ coefficients of kidney and spleen of Juema pigs were significantly lower than Bama miniature pigs (P<0.05 for both).The heart and lung coefficients of Juema pigs were significantly higher than that of Bama miniature pigs (P<0.05 for both).The VEGF and HIF-1α expressions in the lung and kidney in Juema pigs were significantly higher than Bama pigs (P<0.05 or P<0.01).Only the EPO expression in in the lung of Juema pigs was significantly higher than that of the Bama miniature pigs (P<0.05).Conclusions These results indicate that the variation in organ coefficients may be resulted from evolutionary factors such as adaptiveness to environmental physical and energy conditions,pathogens,and energy metabolism demands,etc.in combination.Juema miniature pigs showing a significantly higher expression of hypoxia-related genes than that in Bama minipigs indicate that it has a strong plateau adaptability by higher gene expressions.

Effects of TGF-β stimulation on mouse splenic dendritic cells / 中华微生物学和免疫学杂志

Objective To investigate the mechanisms modulating the functions of dendritic cells (DCs) and suppressing the activation and proliferation of T cells by transforming growth factor-β (TGF-β).Methods Mouse splenic DCs were purified with CD11c+ immunomagnetic beads and the purity of isolated DCs were detected by flow cytometry.Gene chip was used to detect gene expression in DCs after stimulation with TGF-β, and then real-time PCR was performed to analyze the differentially expressed genes in microarray at mRNA level.The activation and proliferation of CD4+ T cells which were co-cultured with DCs after stimulation with TGF-β were detected by flow cytometry.Results The purity of DCs reached over 95% after isolation.TGF-β down-regulated the expression of cell surface markers CD53, CD69, CD33, CD74 and CD93 on DCs;decreased the expression of chemokines Ccl3, Ccl5, Ccl9, Ccl6, Ccl17, Cxcl10, Ccl22, Ccl4, Ccr7, Ccl2, Cxcl9 and Ccl7;inhibited the expression of inflammatory cytokines IL-2ra, IL-12rb2, IL-15ra, IL-1b and IL-15.Moreover, the DCs-mediated activation and proliferation of CD4+ T cells were suppressed by TGF-β.Conclusion TGF-β inhibits the DCs-mediated activation and proliferation of CD4+ T cells by suppressing the expression of surface markers on DCs and down-regulating the expression of chemokines and inflammatory cytokines.

Therapeutic Efficacy of Acupuncture at the Thirteen Ghost Points for Postpartum depression and Its Effect on the Quality of Life / 上海针灸杂志

Objective To observe the therapeutic efficacy of acupuncture at the thirteen ghost points in treating postpartum depression and its effect on the quality of life. Methods Sixty patients with postpartum depression were randomized into a treatment group and a control group, 30 in each group. The treatment group was intervened by acupuncture at the thirteen ghost points, while the control group was by oral administration of Fluoxetine. The Hamilton Rating Scale for Depression (HAMD) and the Short-Form (36) Health Survey (SF-36) were compared before and after intervention. Results The HMAD score was significantly changed in both groups after intervention (P＜0.05). There was a significant difference in comparing the HAMD score between the two groups after intervention (P＜0.05). There were significant differences in comparing the post-treatment bodily pain (BP), general health (GH), vitality (VT), social functioning (SF), mental health (MH), and health transition (HT) between the two groups (P＜0.05). Conclusion Acupuncture at the thirteen ghost points can effectively improve the postpartum depression and the quality of life.

Toxicity of blastula culture medium to mouse embryos: a safety evaluation ofinvitro assisted reproduction devices / 中国组织工程研究

BACKGROUND:The blastula culture medium can assist the development of zygote from the fertilized egg to the blast blastula. The safety and quality of blastula culture medium directly influences the quality of blastula. OBJECTIVE:To evaluate the effect of blastula culture medium on the development of mouse embryos. METHODS: In this study, B6D2F1 mice were used. The female mice were superovulated and mated with male B6D2F1 mice. One day later, the zygotes were colected and cultured in the M16 medium to 4-cel stage. Then, 4-cel stage embryos were transferred into the tested blastula culture medium (experimental group). After 5 days of culture, the forming rate of blastula was examined. Meanwhile, the M16 medium containing endotoxin was used to culture 1-cel mouse zygote (positive control group). The M16 medium with no embryo toxicity was used to culture 1-cel zygote (negative control group). RESULTS AND CONCLUSION:The formation rate of blastula was 0 in the positive group, 87.1% in the negative control group, and 87.3% in the experimental group. From the results, the tested blastula culture medium could assist the 1-cel zygote growing to the stage of blastula, and the formation rate of blastula was above 80%. The tested blastula culture medium had no toxicity to the mouse embryo.

Effect of herbal-partitioned moxibustion at Tianshu (ST 25) and Qihai (CV 6) on pain-related behavior and emotion in rats with chronic inflammatory visceral pain / 针灸推拿医学（英文版）

Objective:To observe the effect of herbal-partitioned moxibustion (HPM) on pain-related behavior and emotion in a rat model of chronic inflammatory visceral pain, and to investigate the mechanism. Methods:Twenty-four male Sprague-Dawley (SD) rats were randomly divided into three groups:a normal group, a model group and an HPM group. Except for the normal group, rats in the other two groups were clystered with mixed liquor of Trinitrobenzene Sulfonic Acid (TNBS) and 50%ethanol to induce the chronic inflammatory visceral pain model. After the models were established successfully, rats in the HPM group were treated with HPM at bilateral Tianshu (ST 25) and Qihai (CV 6). Rats in the normal group and the model group were only fixed as those in the HPM group without treatment. Abdominal withdrawal reflex (AWR) score, mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were adopted to detect the visceral and somatic pain;meanwhile, open field test (OFT) and elevated plus maze test (EPMT) were employed to evaluate pain emotions such as depression and anxiety. Results:Compared with the normal group, AWR scores of the model group were significantly increased under different stimulus expansion pressure level (P Conclusion:HPM has analgesic effect on chronic inflammatory visceral pain. It can reduce the visceral and somatic pain in rats and markedly improve the emotions such as anxiety and depression induced by chronic visceral pain.

Comparison of the Bama minipig and Juema minipig models of high altitude multi-organ dysfunction syndrome / 中国比较医学杂志

Objective To compare the Bama minipig and Juema minipig models of high altitude multi-organ dysfunction syndrome.Methods Six plateau-origin Juema minipigs and plain-origin Bama minpigs in each group received intravenous infusion of 0.35 mg/kg lipopolysaccharide ( LPS) , respectively.Blood samples were taken at 0 h, 3 h, 6 h, 12 h, 24 h, 48 h and 72 h after LPS infusion.Routine blood test was performed, blood CK, AST, ALT, TBIL, CRE were assayed, and histopathological examination of the lung tissues was performed at 24 h, 48 h after LPS infusion.Results The mortality of Bama minipigs was 33.3%, higher than that of 16.7%of Juema minipigs.The trend of physiological and biochemical changes was similar, but was milder in the Juema minipigs than in Bama minipigs.The lung injuries of the Bama minipigs at 24 h and 48 h were more severe than those in the Juema minipigs.Conclusions Both Bama and Juema minipig models of high altitude multi-organ dysfunction syndrome can be successfully established.Juema minipig models can be more closely and safely established, due to its own plateau biological properties, and avoid the influence by extrinsic injurious effects of plateau environment.

The difference and correlation of the expression of hypoxia reaction genes at different time of kainic acidinduced epilepsy in rats / 中华行为医学与脑科学杂志

Objective To investigate the difference of the expression of hypoxia reaction genes of vascular endothelial growth factor(VEGF),erythropoietin (EPO) and hypoxia-inducible factor-1 alpha (HIF-1α)) at different times of rats that were induced epilepsy by kainic acid (KA),and analyze their correlation.Methods The epileptic rat model was established by intraperitoneal injection of kanic acid.The expression of VEGF,EPO and HIF-1α gene were detected by real-time fluorescent quantitative PCR assay with TaqMan probe in different times after intraperitoneal injection of KA.Results Compared with normal sodium (NS) group,the expression VEGF was higher at 12 h((8.38±1.27) ×10-3 ng/μl,P＜0.05)) and 24 h((8.30±5.08) ×10-3 ng/μl,P＜0.05)),EPO was higher at 12 h((8.42±0.90) × 10-5 ng/μl,P＜0.05)) and 48 h ((1.50±3.25) × 10-2 ng/μl,P＜0.01)) while the HIF-1α was higher at 24 h((2.11±0.21) × 10-2 ng/μl,P＜0.01)),48 h((1.50±0.33) × 10-2 ng/μl,P＜0.05))and 72 h((1.64±0.16) × 10-2 ng/μl,P＜0.01)).Furthermore,the expression of EPO showed significant correlations with HIF-1α and VEGF (r=0.573,0.471,P＜0.05),VEGF and HIF-1α had eminent correlations (r=0.803,P＜0.01).Conclusions The expression of VEGF,EPO and HIF-1α participate in the seizure procedure and there is certain correlation between the three genes.

Alpha-Gal antigen and immunity risk control of animal-derived medical devices / 中国组织工程研究

BACKGROUND:Medical devices from animals are commonly used in clinical application. Despite their efficiency is widely accepted, their safety, especialy immunity has been concerned. OBJECTIVE:To investigate immunity risk control to medical devices from animals for safety consideration. METHODS:Using “α-Gal antigen, immunity, xenotransplantation” in Chinese and English as the key words, the first author conducted a computer search of Science direct database (www.sciencedirect.com), Wiley-Blackewel database (http://onlinelibrary.wiley.com) and Wanfang database (www.wanfang.com.cn) through screening the titles and abstracts. RESULTS AND CONCLUSION:Increasing evidence shows that, Gal α1-3Gal antigen (α-Gal antigen) is recognized as the major antigen and abundantly expressed on glycoconjugates of non-primate mammals and New World monkeys. In contrast, the α-gal epitope is not expressed on glycoconjugates of humans and Old World monkeys. Instead, they produce a very large amount of natural anti-α-Gal antibody that specificaly binds the α-gal epitope. The binding of human natural anti-α-Gal to α-gal epitopes expressed on non-primate mammal animals was expected to be unique immunological barrier in xenotransplantation. Therefore, it is important to choose raw materials, reduce or eliminate the α-Ggal epitope, establish highα-Ggal epitope detection methods with high sensitivity and good repeatability for achieving a greater safety and efficiency of medical devices from animals.

Regulation of Moxibustion on the Expression of NF-κBp65 and PPARγ mRNA in Colon of Rats with Ulcerative Colitis / 针灸推拿医学（英文版）

Objective: To explore the mechanism of herb-partitioned moxibustion for ulcerative colitis (UC) through observing the colonic mucosal histopathological changes and the expression of nuclear factor kappaB (NF-kB) and peroxisome proliferators-activated receptorγ (PPARγ) mRNA of UC rats. ＜br> Methods: Male SD rats were randomly divided into a normal group and a model group. UC model was established by general immunological plus local irritation method. After model identification, rats in the model group were randomly divided into a model group, a herb-partitioned moxibustion (HPM) group and a Western medicine (Salicylazosulfapyridine, SASP) group. Rats in the HPM group received treatment at bilateral Tianshu (ST 25) and Dachangshu (BL 25), two cones for each point, once a day for 7 d. SASP group rats were gavaged with SASP. The pathological scores were evaluated according to hematoxylin-eosin (HE) staining of colonic tissues. We used light microscopy to observe degree of colonic mucosal damage and the quantitative polymerase chain reaction (QPCR) to detect the expression of NF-κBp65 and PPARγ in colorectal mucosa. ＜br> Results: Compared with the normal group, histopathological scores were significantly higher in the model group (P＜0.05); compared with the model group, the scores were decreased significantly in the HPM group and the SASP group. Compared with the normal group, NF-κBp65 mRNA expression was increased with statistical significance in the model group (P＜0.05); compared with the model group, NF-κBp65 mRNA expressions were decreased significantly in the HPM group and the SASP group. Compared with the normal group, PPARγ mRNA expression was increased significantly in the model group (P＜0.05); compared with the model group, PPARγ mRNA expressions were decreased significantly in the HPM group and the SASP group. ＜br> Conclusion: HPM could improve the mucosa damage of UC rats, which is possibly through down-regulating NF-κBp65 to achieve anti-inflammatory effect. Whether decreasing the PPARγ mRNA is possibly involved in preventing precancerosis will need further study.

Summary of the Background and Characteristics of Recent-times Shanghai Acupuncture-moxibustion Development / 上海针灸杂志

The historical background of recent-times“developing the Western and restraining the Chinese, and abolishing the old and conserving the new”determines the fact that the prominent feature of acupuncture development is actual effect. Shanghai famous doctors gathered, wrote kooks, established theories, run schools, paid attention to information exchange, participated in treatment for refractory and serious diseases including epidemic diseases, worked together to the academic inheritance of acupuncture and moxibustion and played an important role in preserving traditional Chinese medicine and spreading acupuncture and moxibustion.

Progress in Research on Immunological Effect of Macrophages in Ulcerative Colitis / 胃肠病学

Macrophages( Mφ)play an important role in the regulation of immune reaction. Different kinds of cellular microenvironment influence the phenotypes and function of Mφ and induce different immunological effect. Researches showed that there was a strong correlation between the dysfunction of Mφ and development of ulcerative colitis( UC ). Targeted regulation on immunological activity of Mφ may improve the clinical manifestations and pathological changes in UC. Regulation of the activity and migration of Mφ through changing microenvironment might be one of the potential mechanisms of moxibustion in treating UC. Progress in research on immunological effect of Mφ in UC was summarized in this review article.

The preparation and bioactivity assessment of gene recombinant fibrin-binding brain derived neurotrophic factor / 生物医学工程学杂志

Using the E. coli, we fabricated the gene reconstructed brain derived neurotrophic factor with a fibrin binding domain (FBD-BDNF). We then tested the neurotrophic bioactivity and fibrin-binding ability of the FBD-BDNF. The E. coli was used to express the recombinant protein. The inclusion body was purified with column chromatography and renaturated to construct the right 3D formation. In this study, we successfully fabricated the FBD-BDNF and tested the binding ability and neurotrophic activity. The results demonstrated that FBD-BDNF had special binding ability of fibrin and significant neurotrophic activity for DRG cells. FBD-BDNF could have a promising application prospect in nerve tissue engineering.

Correlation between peripheral blood inflammatory cytokines and kainic acid-induced seizure severity in rats / 中华行为医学与脑科学杂志

Objectives To investigate the correlation between the level of peripheral blood inflammatory cytokines and kainic acid-induced seizure severity in rats.Methods 140 rats were divided into control and model group randomly,70 rats in each group.Model group rats were injected with kainic acid (10 mg/kg) by intraperitoneal,and the control rats were injected with sodium chloride.The change of their behaviors was observed and the concentrations of TNF-α,IL-1β,IL-4 and IL-10 were determined by ELISA in each group at different times.Results The rats showed epilepsy grand mal in 3 h-9 h after KA injection.The concentrations of TNF-α and IL-1β in model groups were significantly higher than those in control group in 6 h-12 h (6 h:(21.5±3.2) pg/ml vs (12.3±3.1)pg/ml;12 h:(20.6±4.2)pg/ml vs (11.5±3.8)pg/ml)(P＜0.05) and IL-4 in model group was higher at only 12 h ((53.55±3.08) pg/ml vs (33.26±4.16)pg/ml) (P＜0.05).The level of IL-10 in model groups was not statistically significant compared with control group (P＞0.05).Conclusion The proinflammatory cytokines (TNF-α and IL-l β) participate in the seizure procedure,meanwhile their levels and seizure severity have eminent correlations,but antiinflammatory cytokines (IL-4 and IL-10) had not.

Research Advance on Moxa Smoke / 针灸推拿医学（英文版）

The purpose was to investigate the research achievements over the past two decades on moxa smoke produced in the process of moxibustion, in terms of its clinical studies, chemical compositions, safety assessment as well as the mild smoke or smoke-free moxibustion, for revealing the recent dynamic and developing orientation, and promoting its further application, succession and innovation.

Recent research and progress of medical dressings / 中国组织工程研究

BACKGROUND: Medical dressings can play a temporality barrier function as skin substitute in wound healing, which can avoid or control wound infection. With the increasing of aging and chronic ulcer wound, medical dressings play a more important role. OBJECTIVE: To review the recent research and progress of medical dressings, in addition, to explore its developing direction. METHODS: Elsevier database and CNKI was retrieved by computer with key words of "medical dressing, collagen, gel and chitosan" to search papers published between January 1980 and January 2009. Related papers addressing medical dressings were selected. According to inclusion cdteda, 35 literatures were selected Jn this study. RESULTS AND CONCLUSION: Currently, the medical dressings can be classified into natural polymer, synthetic macromolecule, inorganic material and composite. Their performances and clinical application were reviewed respectively. The quality control and future development of medical dressing products were also discussed. This paper can provide a theoretical foundation for the researcher in study and development of medical dressings, manufacturer in the quality control and government in product quality supervision.

Detection of unknown residual solvents of sodium hyaluronate product / 中国组织工程研究

BACKGROUND: A mass of unknown remains are founded in sodium hyaluronate product when it was tested for quality control. OBJECTIVE: To qualify and quantify unknown residual solvents of sodium hyaluronate product and determine hazardsaccording to standard toxicological data. DESIGN, TIME AND SETTING: A quality and quantity study with combined gas chromatography mass spectrometry was performed at National Institute for the Control of Pharmaceutical and Biological Products from May to June 2007.MATERIALS: Experimental samples were spot-checked, and purified water was also used.METHODS: The samples were qualified and quantified using combined gas chromatography mass spectrometry.MAIN OUTCOME MEASURES: Methanol, xylene and ethyl benzene were qualified and quantified.RESULTS: Combined gas chromatography mass spectrometry demonstrated that residual solvents in the sodium hyaluronateproducts were methanol, xylene, and ethyl benzene. The quantization of methanol was 414.365 μg/mL, the quantization of o-xylene was 0.19 μg/mL, and the quantization of ethyl benzene was 0.22 μg/mL. CONCLUSION: Methanol, xylene, and ethyl benzene were firstly identified as residual solvents in sodium hyaluronate products. Besides, we discussed methods of qualifying and quantifying these three residual solvents.