Bilineage embryo-like structure from EPS cells can produce live mice with tetraploid trophectoderm

Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro.

Advances in anti-adversity of biological composites based on metal-organic frameworks / 生物工程学报

Metal-organic frameworks (MOFs) are formed by self-assembly of metal ions or clusters with organic ligands, and are widely used in the fields of catalysis, sensing, energy and biomedicine. Recently, biological composites based on MOFs have attracted increasing attention. MOFs can be used as a platform for encapsulating bioactive substances due to the advantages such as large pore capacity, large specific surface area and diverse structure composition. These features can protect bioactive substances from adverse conditions, e.g. high temperature, high pressure, and organic solvents, thus improving the anti-adversity of bioactive substances. This review summarizes the advances of using MOFs as protective coatings to improve the anti-adversity of different bioactive substances, and introduces the synthesis strategy of MOFs-based biological composites, with the aim to promote the practical application of MOFs-based biological composites.

Fabrication and biocompatibility assessment of the scaffold with biomimetic interconnected macropore structure / 中国组织工程研究

BACKGROUND: Development of bone tissue engineering provides a new method to solve bone defect repair. Scaffold material and structure construction are issues of concern. OBJECTIVE: To fabricate a silk fibroin and hydroxyapatite scaffold with biomimetic interconnected macropores structure using the paraffin-sphere leaching technology and to evaluate its possibility of cytocompatibility. METHODS: The scaffold with biomimetic interconnected macropores structure was made by the paraffin-sphere leaching technology. The structure of scaffold was observed by the stereomicroscope and scanning electron microscope. The pore size and elasticity modulus were calculated. Passage 3 rabbit adipose-derived mesenchymal stem cells were seeded into the scaffold. The cell viability was detected by live/dead staining at 48 hours after culture. The cell adhesion was observed by hematoxylin-eosin staining at 1 week of culture. The scaffold-cell composite was observed under scanning electron microscope at 3 days of culture. The cell proliferation was detected by the cell counting-kit 8 assay at 1, 3, 5 and 7 days of culture. Those cells cultured alone were considered as control group. RESULTS AND CONCLUSION: (1) Stereomicroscope showed the ivory silk fibroin/hydroxyapatite scaffold. Scanning electron microscope revealed pore structures in cross-section and longitudinal-section with good connectivity. The scaffold pore size was (362.23±26.52) μm and the elasticity modulus was (54.93±5.44) kPa. (2) Scanning electron microscope showed that adipose-derived mesenchymal stem cells adhered and stretched well in the pore wall and connected pore, secreted abundant extracellular matrix, and filled in the pores of silk fibroin/hydroxyapatite scaffold. (3) Hematoxylin-eosin staining results found that adipose-derived mesenchymal stem cells evenly adhered onto the inner wall of silk fibroin/hydroxyapatite scaffold, and proliferated well. (4) Live/dead staining revealed a good viability of adipose-derived mesenchymal stem cells. (5) Cell counting-kit 8 assay results showed the good proliferation of adipose-derived mesenchymal stem cells on the scaffold. (6) To conclude, the silk fibroin/hydroxyapatite scaffold possesses good pore size and cytocompatibility.

In vitro and in vivo effects of 5-aminolevulinic acid-mediated photodynamic therapy against oral squamous cell carcinoma / 中华口腔医学杂志

Objective@#To investigate the in vitro and in vivo effects of 5-aminolevulinic acid (5-ALA)-mediated photodynamic therapy against oral squamous cell carcinoma (OSCC) and preliminarily explore the possible mechanisms.@*Methods@#SCC25 cells were divided into the control group (5-ALA of 0 mg/L) and the experimental group (5-ALA of 10, 25, 50, 100 and 150 mg/L). The production of protoporphyrin Ⅸ (PpⅨ) induced by 5-ALA in SCC25 cells was detected using the flow cytometry. SCC25 cells were divided into the control group (5-ALA of 0 mg/L), lazer alone group, 5-ALA alone group (5-ALA of 100 mg/L) and the 5-ALA combined with laser irradiation group (5-ALA of 5, 10, 25, 50 and 100 mg/L), the cytotoxicity of 5-ALA combined with laser irradiation (wave length 635 nm, power density 87 mW/cm2 and laser dose 10.4 J/cm2) was evaluated in SCC25 cells using the methyl thiazolyltetrazolium assay (incubation times of 4, 8 and 12 h in each group) and the induction effect of combination treatment on the cell apoptosis was assessed by the flow cytometry (incubation time of 12 h in each group). The intracellular production of reactive oxygen species (ROS) triggered by 5-ALA combined with laser irradiation was determined using a fluorescence probe method (incubation time of 12 h in each group). A mouse OSCC xenograft model bearing SCC25 tumor was built, and the mice were divided into control group (saline), 5-ALA group (5-ALA of 50 mg/kg) and 5-ALA combined with laser irradiation group (5-ALA of 10, 25 and 50 mg/kg). Antitumor effect of 5-ALA combined with laser irradiation (wave length 635 nm, power density 158 mW/cm2 and laser dose 94.8 J/cm2) was further measured.@*Results@#5-ALA induced the production of PpⅨ in SCC25 cells in a drug concentration (0-150 mg/L)-and incubation time (0-24 h)-dependent manner. When the 5-ALA concentration was 100 mg/L, the intracellular PpⅨ production was in a relatively stable state. Cell viability and apoptosis rate of 5, 10, 25, 50, 100 mg/L 5-ALA combined with laser irradiation are, respectively, (82.3±5.2)%, (3.13±0.38)%; (74.6±9.3)%, (5.38±0.55)%; (38.3±9.7)%, (17.97±2.72)%; (9.2±3.8)%, (24.47±3.37)%; (7.2±0.8)%, (43.01±5.96)%, which indicated that 5-ALA combined with laser irradiation notably inhibited the growth of SCC25 cells and also induced significant cell apoptosis compared with the control group [(96.3±6.0)%, (0.35±0.13)%, P<0.05]. After combination treatment (5-ALA of 5, 10, 25, 50 and 100 mg/L combined with laser irradiation, the mean fluorescence intensity of dichlorofluorescein is (1.46±0.12)×104, (2.16±0.30)×104, (3.57±0.34)×104, (81.70±13.05)×104, (113.00±7.35)×104, respectively, a large amount of ROS was produced in SCC25 cells compared with the control group [(0.96±0.15) ×104, P<0.05], which was in positive correlation with the intracellular PpⅨ content. 5-ALA (concentration of 10, 25 and 50 mg/kg) combined with laser irradiation greatly suppressed the tumor growth in SCC25 tumor-bearing mice compared to the control group (P<0.05).@*Conclusions@#5-ALA-mediated photodynamic therapy can trigger the generation of intracellular ROS that has significant cytotoxicity and apoptosis induction effect, and thus inhibit the tumor growth both in vitro and in vivo.

[Preparation and characterization of oriented scaffolds derived from cartilage extracellular matrix and silk fibroin].

OBJECTIVE: This study aims to prepare oriented scaffolds derived from a cartilage extracellular matrix (CECM) and silk fibroin (SF) and use to investigate their physicochemical property in cartilage tissue engineering. METHODS: Oriented SF-CECM scaffolds were prepared from 6% mixed slurry (CECM：SF=1：1) through modified temperature gradient-guided thermal-induced phase separation, followed by freeze drying. The SF-CECM scaffolds were evaluated by scanning electron microscopy (SEM) and histological staining analyses and determination of porosity, water absorption, and compressive elastic modulus of the materials. RESULTS: The SEM image showed that the SF-CECM scaffolds contained homogeneous reticular porous structures in the cross-section and vertical tubular structures in the longitudinal sections. Histological staining showed that cells were completely removed, and the hybrid scaffolds retained proteogly can and collagen. The composition of the scaffold was similar to that of natural cartilage. The porosity, water absorption rate, and vertical compressive elastic modulus of the scaffolds were 95.733%±1.010%, 94.309%±1.302%, and (65.40±4.09) kPa, respectively. CONCLUSIONS: The fabricated SF-CECM scaffolds exhibit satisfactory physicochemical and biomechanical properties and thus could be an ideal scaffold in cartilage tissue engineering.

Application of 70 kV tube voltage combined with low dose and low concentration of contrast medium in CCTA / 实用放射学杂志

Objective To access the effects of 70 kV tube voltage combined with low dose and low concentration of contrast medium in coronary CT angiography (CCTA)by evaluating the image quality,radiation dose and contrast medium dosage.Methods Ninety patients suspected with coronary artery disease with body mass index(BMI)of less than 25 kg/m2and heart rate (HR)of less than 75 beats per minute were enrolled.The patients were randomly divided into three groups (n=30 for each group):Group A,100 kV of tube voltage with 370 mg I/mL iopromide,1 mL/kg;Group B,80 kV with 270 mg I/mL iodixanol,1 mL/kg;Group C,70 kV with 270 mg I/mL iodixanol,0.8 mL/kg.All the patients underwent CCTA with a 256 row wide-coverage volumetric CT.Automatic tube current modulation technique was applied.The images were reconstructed by adaptive statistical iterative reconstruction V (ASIR-V).The subjective image quality scores were compared with rank-sum test.The signal-to-noise ratio (SNR),contrast-to-noise ratio(CNR),effective dose(ED)and the total iodine intake were calculated and compared with one-way ANOVA.Results No statistically differences in age,gender,heart rate and BMI were observed among the three groups (P>0.05).Subjective image quality scores had no difference among the three groups (P>0.05).The CT values of group C were higher than those of group A(P<0.05).The image noise of group C was higher than that of group A and group B (P<0.05).No significant differences in SNR and CNR were noticed among the three groups (P>0.05).The ED of group B (0.39±0.08)mSv and group C (0.19±0.01)mSv were lower than that of group A (0.81±0.19)mSv (each P<0.05).Compared with group A,the decrease rates of ED of group B and C were 51.8% and 76.5% respectively.Compared with group A and group B,the total iodine intake of group C was decreased by 25% and 21.4% (P<0.05).Conclusion 70 kV of tube voltage combined with low dose and low concentration of contrast medium in CCTA can reduce the radiation dose and iodine intake without compromising image quality.

Osteochondral tissue engineering based on biomimetic osteochondral scaffold contained calcified cartilage layer compounding with ADSCs in vivo / 中华骨科杂志

Objective The biomimetic osteochondral scaffold contained calcified cartilage layer(CCL) was fabricated using slik fibroin (SF) and hydroxyapatite (HA) for materials.To investigate effects of biomimetic osteochondral scaffold contained CCL compounding with ADSCs on regeneration of the osteochondral defect on the rabbit knee,explore the feasibility of this design as a concept of osteochondral tissue engineering.Methods We fabricated a novel biomimetic osteochondral scaffold with CCL using SF and HA by the combination of paraffin-sphere leaching and modified temperature gradient-guided thermal-induced phase separation (TIPS) technique.The pore size,porosity,and compressive modulus of elasticity of the scaffold cartilage layer and the osteogenic layer were measured by scanning electron microscopy and microscopy CT.The osteochondral defect model on rabbit bilateral knees were established,and implanted with the non-CCL group (non-CCL scaffold compounding with ADSCs) and CCL group (CCL scaffold compounding with ADSCs).At 4,8 and 12 weeks after implantation,the rabbits were euthanized,respectively.Gross observation score,histological and immunohistochemical assessment,biochemical quantitative of new osteochondral tissue,micro-CT scans for new bone,were executed.We evaluated the regeneration of osteochondral defects in each group,and verified the role of CCL in vivo.Results The biomimetic osteochondral scaffold with CCL had a consecutively overlapping trilayer structure with different densities and pore structures,including a chondral layer (top layer),intermediate layer and bony layer (bottom layer).The cartilage layer had a well-oriented microporous structure with a uniform distribution with a pore size of (112.43± 12.65)μm and a porosity of 90.25％±2.05％.The subchondral bone layer had a good three-dimensional macroporous structure,good connectivity,pore size (362.23±26.52) μm,porosity of osteogenic layer was 85.30％± 1.80％.The cartilage regeneration in CCL+AD-SCs group was better than non-CCL+ADSCs group.The content of GAG and type Ⅱ collagen in new cartilage tissue in CCL+AD-SCs group was much more than non-CCL+ADSCs group.The new bone tissue analysis and biomechanical testing had no significant differences between the two groups.Conclusion The biomimetic osteochondral SF/HA scaffold contained CCL mimics the structure of normal osteochondral tissue with good 3-dimensional pore structure and biocompatibility.The scaffold complex autologous ADSCs successfully repair osteochondral defects in rabbit knee,and the presence of CCL accelerates the growth of cartilage.

Protective effect of HSP27 on Hcy induced vascular endothelial cell injury and its mechanism / 重庆医学

Objective To study the protective effect of heat shock protein 27(HSP27)on homocysteine(Hcy)induced vascular endothelial cell injury and its mechanism.Methods Human umbilical vein endothelial cells(HUVECs) were cultured.The influence of Hcy on the survival rate of HUVECs was detected by MTT method;the Annexin V FITC apoptosis detection kit and flow cytometry were used to detect the influence of Hcy and HSP27 on cellular apoptosis;the experiment design adopted the total nitric oxide detection kit(nitrate reductase method) for detecting the NO level in cell culture fluid,the DCFH DA reactive oxygen species (ROS) detection kit and flow cytometry were used to detect intracellular ROS to research possible protection mechanism.Results After action of HUVECs with different concentrations of Hcy,the mean values of survival rates in the 0 mmol/L,0.2 mmol/L,0.4 mmol/L,0.6 mmol/L,0.8 mmol/L and 1.0 mmol/L groups were 97.33％,95.17％,90.72％,78.29％,63.65％ and 41.51％ respectively.The difference between the 0 mmol/L group and 0.2 mmol/L group had no statistical significance(P＞0.05);the survival rate in the Hcy 0.4,0.6,0.8,1.0 mmol/L groups was lower than that in the 0 mmol/L group,the difference was statistically significant(P＜0.05).The cellular survival rate was negatively correlated with the Hcy concentration.The higher the HSP27 concentration,the higher the cellular survival rate,showing the concentration dependence.HSP27 had a protective effect on HUVECs damage.Hcy could induce apoptosis of endothelial cells,while HSP27 could significantly inhibit apoptosis,indicating that HSP27 could reduce the damage of Hcy on endothelial cells.HSP27 could inhibit the NO decrease caused by Hcy and inhibited the production of Hcy induced ROS.Conclusion HSP27 protects Hcyinduced injured endothelial cells by influencing NO expression and regulating intracellular ROS level.

Effects of serine/threonine-protein kinase B-Raf-activated long-chain non-coding RNA on apoptosis and autophagy in thyroid carcinoma cells / 中南大学学报(医学版)

To investigate the effect of serine/threonine-protein kinase B-Raf (BRAF)-activated long-chain non-coding RNA (lncRNA-BANCR) on apoptosis and autophagy in thyroid carcinoma cells and the underlying mechanisms.
 Methods: RT-PCR was used to detect the expression of lncRNA-BANCR in thyroid carcinoma and normal thyroid tissues. The association between lncRNA-BANCR and clinicopathological data was analyzed in patients with thyroid cancer. Cell counting kit-8 (CCK-8) was used to detect the effect of lncRNA-BANCR on the proliferation of thyroid cancer cells. The effect of lncRNA-BANCR on the apoptosis of thyroid carcinoma cells was detected by flow cytometry. Transwell invasion assay was used to detect the effect of lncRNA-BANCR on the invasive ability of thyroid cancer cells. Western blot was used to detect the changes of autophagy proteins LC3-I and LC3-II after the lncRNA-BANCR expression was suppressed.
 Results: Compared with normal thyroid tissues, the expression level of lncRNA-BANCR in thyroid carcinoma tissues was elevated (P<0.05). The expression of lncRNA-BANCR was positively related to the pathological stage of thyroid carcinoma and the lymph node metastasis. Inhibition of lncRNA-BANCR expression attenuated the proliferation and invasion ability of thyroid cancer cells (both P<0.05); but the apoptosis was enhanced (P<0.05); the expression levels of autophagy protein LC3-I and LC3-II were also increased (P<0.05).
 Conclusion: The expression level of lncRNA-BANCR affects the proliferation, invasion and apoptosis of thyroid cancer cells through modulation of autophagy behavior.

[Development and characterization of oriented scaffolds derived from cartilage extracellular matrix].

OBJECTIVE: This study aimed to prepare oriented scaffolds derived from a cartilage extracellular matrix (CECM) and to investigate their physicochemical property and compatibility with adipose-derived stem cells (ADSCs). METHODS: A fresh porcine articular cartilage was cut into pieces. Cartilage nanofibers with diameters of 50-500 nm were collected through homogenization and centrifugation. These nanofibers were then decellularized by using Triton X-100 to produce 6% CECM. The oriented scaffolds derived from the nanoscale CECM were fabricated through unidirectional solidification and lyophilization. Afterward, these scaffolds were crosslinked. The physical and chemical performances and cell compatibility of CECM-oriented scaffolds were evaluated. RESULTS: The cross-sections of the scaffolds contained homogeneous reticular porous structures with nanofibers on the walls of the pores, and the longitudinal sections revealed vertical tubular structures. Hematoxylin-eosin staining revealed that the scaffolds were red without blue. Toluidine blue, safranin O, and Sirius red staining showed positive results. The porosity, water absorption rate, and vertical compressive elastic modulus of the scaffolds were 95.455%±0.910%, 95.889%±1.071%, and (40.208±5.097) kPa, respectively. CONCLUSIONS: The components of the oriented scaffolds derived from CECM are similar to those of native cartilage with favorable biocompatibility. The porous structures and sizes of the scaffolds are suitable for the adhesion, proliferation, and infiltration of ADSCs. The oriented scaffolds derived from CECM are relatively optimal for cartilage tissue engineering. 
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The research progress in transfusion-related iron overload in hematological malignancies / 实用肿瘤学杂志

Transfusion -related iron overload damage iron homeostasis of organism ,which damages the structure of the vital organs and leads to the dysfunction of important organs .Iron overload affects the prognosis of the patients,which promotes myelodysplastic syndrome transformation to acute leukemia and reduces the total sur -vival rate of patients with hematopoietic stem cell transplantation ,which increases the treatment related mortality after hematopoietic stem cell transplantation and increases the chance of infection .Iron depletion therapy can de-lay the progression of acute leukemia and prolong the survival time of the patients with acute leukemia and show a certain anti-proliferative activity .

Development and characterization of oriented scaffolds derived from cartilage extracellular matrix / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aimed to prepare oriented scaffolds derived from a cartilage extracellular matrix (CECM) and to investigate their physicochemical property and compatibility with adipose-derived stem cells (ADSCs).</p><p><b>METHODS</b>A fresh porcine articular cartilage was cut into pieces. Cartilage nanofibers with diameters of 50-500 nm were collected through homogenization and centrifugation. These nanofibers were then decellularized by using Triton X-100 to produce 6% CECM. The oriented scaffolds derived from the nanoscale CECM were fabricated through unidirectional solidification and lyophilization. Afterward, these scaffolds were crosslinked. The physical and chemical performances and cell compatibility of CECM-oriented scaffolds were evaluated.</p><p><b>RESULTS</b>The cross-sections of the scaffolds contained homogeneous reticular porous structures with nanofibers on the walls of the pores, and the longitudinal sections revealed vertical tubular structures. Hematoxylin-eosin staining revealed that the scaffolds were red without blue. Toluidine blue, safranin O, and Sirius red staining showed positive results. The porosity, water absorption rate, and vertical compressive elastic modulus of the scaffolds were 95.455%±0.910%, 95.889%±1.071%, and (40.208±5.097) kPa, respectively.</p><p><b>CONCLUSIONS</b>The components of the oriented scaffolds derived from CECM are similar to those of native cartilage with favorable biocompatibility. The porous structures and sizes of the scaffolds are suitable for the adhesion, proliferation, and infiltration of ADSCs. The oriented scaffolds derived from CECM are relatively optimal for cartilage tissue engineering. 
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[Fabrication and evaluation of hydroxyapatite-chitosan scaffold via simulated body fluid biomimetic mineralization].

OBJECTIVE: This research aimed to fabricate a hydroxyapatite (HA)-chitosan scaffold via simulated body fluid (SBF) biomimetic mineralization and determine how mineralization time affects scaffold construction and cell compatibility. METHODS: The HA-chitosan scaffolds were fabricated by freeze-drying technique and then subjected to precalcification, also known as alternative soaking method. Afterward, precalcificated scaffolds were placed into the SBF to conduct the mineralization process. Mineralization time was set at 7, 14, and 21 days, corresponding to three experimental groups. Pure chitosan scaffolds acted as the control group, and the physical and chemical properties of the four groups were tested. Osteogenic-induced adipose-derived stem cells (ADSCs) were seeded into the scaffolds to investigate the scaffolds' cell compatibility. RESULTS: The mineral substance of the 14-day group exhibited a uniform distribution. The crystal composition of the mineral substance suited the HA's features. The compressive elastic modulus increased along with the extension of mineralization time. The 21-day group showed a statistically significant increase in compressive elastic modulus compared with the control group (P < 0.05). The cell proliferation level of the 14-day group was significantly the highest among the three experimental groups (P < 0.05). The calcium ion and the type I collagen had the highest secretion amount when the cells were seeded into the 14-day group. CONCLUSION: The SBF biomimetic mineralization method can be used to fabricate HA-chitosan bone-tissue-engineering scaffolds. The biological compatibility, as well as the chemical and physical properties, reached the optimum levels at day 14.

Application of the PD-1 antibody in melanoma therapy / 中国肿瘤临床

A melanoma is a highly malignant tumor that originates from melanocytes in the skin, mucosa, or tunica pigmentosa. The incidence and mortality rate of cutaneous melanoma are increasing annually. However, the efficacy of traditional therapy is extremely limited because of its low sensitivity and high toxicity. The application of the anti-CTLA-4 antibody and the BRAF inhibitor dramatically improves the overall survival of patients with advanced melanoma. However, their limited benefit ratio and high drug resistance curtail the use of anti-CTLA-4. Since the US Food and Drug Administration approved the use of the anti-PD-1 and anti-PD-L1 antibodies for ad-vanced melanoma in 2014, a significant survival benefit has been observed in patients with advanced melanoma. This review aims to highlight the applications of the anti-PD-1 antibody (pembrolizumab, nivolumab, and pidilizumab) and the anti-PD-L1 antibody (MP-DL3280A, BMS-936559, and MEDI4736) in the clinical treatment of melanoma by succinctly summarizing the results of recent reports.

Fabrication and evaluation of hydroxyapatite-chitosan scaffold via simulated body fluid biomimetic mineralization / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This research aimed to fabricate a hydroxyapatite (HA)-chitosan scaffold via simulated body fluid (SBF) biomimetic mineralization and determine how mineralization time affects scaffold construction and cell compatibility.</p><p><b>METHODS</b>The HA-chitosan scaffolds were fabricated by freeze-drying technique and then subjected to precalcification, also known as alternative soaking method. Afterward, precalcificated scaffolds were placed into the SBF to conduct the mineralization process. Mineralization time was set at 7, 14, and 21 days, corresponding to three experimental groups. Pure chitosan scaffolds acted as the control group, and the physical and chemical properties of the four groups were tested. Osteogenic-induced adipose-derived stem cells (ADSCs) were seeded into the scaffolds to investigate the scaffolds' cell compatibility.</p><p><b>RESULTS</b>The mineral substance of the 14-day group exhibited a uniform distribution. The crystal composition of the mineral substance suited the HA's features. The compressive elastic modulus increased along with the extension of mineralization time. The 21-day group showed a statistically significant increase in compressive elastic modulus compared with the control group (P < 0.05). The cell proliferation level of the 14-day group was significantly the highest among the three experimental groups (P < 0.05). The calcium ion and the type I collagen had the highest secretion amount when the cells were seeded into the 14-day group.</p><p><b>CONCLUSION</b>The SBF biomimetic mineralization method can be used to fabricate HA-chitosan bone-tissue-engineering scaffolds. The biological compatibility, as well as the chemical and physical properties, reached the optimum levels at day 14.</p>

Regulating Effect of Acupuncture at Fengchi (GB20) and Extraordinary Points on TNF-?and ET in Hypertension / 上海针灸杂志

ObjectiveToobserve the regulating effect of acupuncture at Fengchi (GB20) and extraordinary points on tumor necrosis factor (TNF)-aand endothelin (ET) in hypertension.MethodTotally 150 patients with hypertension were randomized into two groups at a ratio of 1:1.In the control group, 75 subjects were intervened by acupuncture at bilateral Fengchi; in the treatment group, 75 subjects were intervened by acupuncture at bilateral Fengchi plus extraordinary points. The needles were manipulated once every 10 min duringthe 30-min session. The treatment was given once per day, 2 weeks as a treatment course. ResultDuring the intervention, the TNF-aand ET contents were decreased significantly in the treatment group (P0.05). After 1 treatment course, the TNF-aand ET contents were decreased significantly in the treatment group (P0.05); there was a significant difference in comparing the TNF-aand ET contents between the two groups (P<0.05). There were also significant differences in comparing the TNF-aand ET contents inhypertension of various degrees between the two groups (P<0.01). The results showed that the TNF-aand ET contents in patients intervened by acupuncture at Fengchiand extraordinary points were lower than that in those intervened by acupuncture at Fengchialone, i.e. combination of Fengchi and extraordinary points produces a significant effect on regulating TNF-aand ET contents.Conclusion Acupuncture at Fengchi and extraordinary points can produce a more significant effect than acupuncture at Fengchi alone on regulating TNF-aand ET in patients with hypertension.

Fabrication and biocompatibility assessment of a silk fibroin scaffold with biomimetic oriented microstructure for cartilage tissue engineering / 天津医药

Objective To fabricate a silk fibroin scaffold with biomimetic oriented microstructure using the directional crystallization technology, and to evaluate the possibility of application to the cartilage tissue engineering. Methods The silk fibroin scaffold with biomimetic oriented microstructure was made by the directional crystallization technology. The structure of scaffold was observed by the scanning electron microscope, and the pore size, porosity and mechanical properties were calculated. Adipose-derived stem cells were isolated from rabbit, and the passage 3 was seeded into the scaffold. The cell proliferation was detected by CCK8 method. The cell adhesion ability was observed by HE staining and scanning electron microscope. The cell viability was observed by LIVE/DEAD staining. Results The scanning electron microscopy showed that the parallel microtubule-like structure can be seen arranged in longitudinal section of the scaffold, which had a uniform directivity, and also the elliptical or circular pore structure in cross-section. The scaffold had a good pore interconnectivity with (112.43±12.65) μm pore diameter of the cross-section, (90.25±2.05)%porosity and (52.48±5.78) kPa the compression modulus. HE staining and scanning electron microscopy demonstrated that the cells uniformly adhered to the surface and inner pore, and which secreted lots extracellular matrix distributed in the oriented microstructure. Results of CCK8 showed that the good cell proliferation on the scaffold, and the LIVE/DEAD staining indicated that the cells were maintained good viability. Conclusion The silk fibroin scaffolds have a biomimetic oriented microstructure, and show good pore diameter, porosity, biocompatibility and biomechanical properties, which made it a suitable candidate as an alternative scaffold for cartilage tissue engineering.

Inhibitory effect of von Willebrand factor-cleaving protease on angiogenesis / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of von Willebrand factor-cleaving protease, a disintegrin-like and metalloprotease with thrombospondin-1 repeats (ADAMTS13)on angiogenesis induced by vascular endothelial growth factor (VEGF)in vitro and in vivo.</p><p><b>METHODS</b>Cell proliferation assay, differentiation (tube formation)assay and wound migration assay were performed by using human umbilical vein endothelial cells (HUVECs)to explore the effect of ADAMTS13 on angiogenesis in vitro. Cells were treated with different concentrations of ADAMTS13 (1, 5, 25, 50 and 100 nmol/L)and the number of cells was counted via MTT assay. In addition, effect of ADAMTS13 on differentiation was assessed by measuring the length of capillary-like tube structures formed by HUVECS in matrigel. Effect of ADAMTS13 on HUVEC migration was assessed via calculation of wound healing distance after 8 hrs culture with VEGF or ADAMTS13. Chick embryo chorioallantoic membrane (CAM) assay and Matrigel plug assay were performed to investigate the effect of ADAMTS13 on angiogenesis in vivo.</p><p><b>RESULTS</b>ADAMTS13 significantly inhibited the proliferation of HUVECs induced by VEGF in a dose-dependent manner. Migration distance of HUVECs was (79 ± 22) μm in control group, (250 ± 8)μm in VEGF-treated group and (170 ± 23)μm in VEGF and ADAMTS13 cotreated-group after 8 hrs, respectively. The tube length is (450.6 ± 16.6)% in VEGF-treated group and (235.3 ± 19.0)% in VEGF and ADAMTS13 cotreated-group of that of control group after HUVECs cultured in matrigel for 16 hrs. The number of blood vessels decreased after treatment with ADAMTS13 in CAM assay. The number of blood vessels was (228.2 ± 10.8)%, (69.2 ± 21.1)%, (184.6 ± 15.2)% in VEGF treated-group, ADAMTS13 treated-group and VEGF and ADAMTS13 cotreated-group of that of control group, respectively. Formation of capillary-like network in matrigel plugs containing VEGF was reduced to 43.5% by ADAMTS13 in matrigel plug assay in mouse model.</p><p><b>CONCLUSION</b>ADAMTS13 inhibits the HUVECs proliferation, differentiation and migration in vitro. ADAMTS13 inhibits chick embryos vascularization and formation of capillary-like network in vivo.</p>

Effect of different durations of low-concentration isoflurane anesthesia on expression of NR2B in hippocampus of mice / 中华麻醉学杂志

Objective To investigate the effect of different durations of low-concentration isoflurane anesthesia on the expression of 2B subunits-containing N-methyl-D-aspartate receptors (NR2B) in the hippocampus of mice.Methods Forty C57BL/6 mice, aged 10 weeks, weighing 20-24 g, were randomly divided into 5 groups (n=8 each) using a random number table: control group (group C);0.7％-1.0％ isoflurane inhaled for 30 min group (group I1);0.7％-1.0％ isoflurane inhaled for 2 h group (group I2);0.7％-1.0％ isoflurane inhaled for 4 h group (group I3);0.7％-1.0％ isoflurane inhaled for 6 h group (group I4).Morris water maze test was carried out at 48 h after the end of anesthesia, and the escape latency and frequency of crossing the original platform were recorded.The mice were sacrificed after the end of the test, and the hippocampi were removed for determination of the expression of activated caspase-3 and NR2B (using Western blot analysis).Results Compared with group C, the escape latency was significantly shortened, and the frequency of crossing the original platform was increased in I1 and I2 groups, no significant changes were found in the activated caspase-3 expression, and the expression of NR2B was up-regulated in I1, I2 and I3 groups, and the escape latency was prolonged, the frequency of crossing the original platform was decreased, the activated caspase-3 expression was up-regulated, and the expression of NR2B was down-regulated in group I4.Conclusion The mechanism underlying the effect produced by different durations of low-concentration isoflurane anesthesia on the cognitive function is related to the expression of NR2B in the hippocampus of mice.