BACKGROUND:Bone mesenchymal stem cells(BMSCs) can accelerate vascularization of myocardial cell,relieve myocardial remodeling,and improve heart function.However,whether its paracrine action can facilitate the differentiation of CD117+ cardiac stem cells(CSCs) by effecting transforming growth factor-? Ⅲ(TGF-? Ⅲ) receptor expression is still unknown.OBJECTIVE:To study the influence of BMSCs on the expression of TGF-? Ⅲ receptor in the process of differentiation of CD117+ CSCs.DESIGN,TIME AND SETTING:In vitro cytology contract observation.The experiment was performed at the Xinhua Hospital from February 2008 to February 2009.MATERIALS:Neonatal rats,weighing 5-8 g,were used to prepare CSCs.Sprague Dawley rats,4 weeks old,weighing 200-250 g,were prepared for BMSCs.METHODS:CD117+CSCs were obtained from neonatal rats by explant cultures and magnetic bead cell sorting.Immunohistochemical experiment was performed to identify specificity of CSCs.CD117+CSCs suspension were synchronization without serum for 24 hours,washed by PBS for 3 times,and then cultured with DMEM/F12,followed by inoculation into 6-well plate.The cells were divided into the control and co-culture groups.In the control group,CSCs were induced differentiation by cardiac spheres,and Transwell chamber were used in the co-culture group.MAIN OUTCOME MEASURES:The growth of CSCs was observed by inverted microscope.The changes of cardiac Troponin T(cTnT),connexin43,TGF-? Ⅲ receptor,smad2 and phospho-smad2 expressions were detected by western blot at days 1,3,5 and 7 after culture.RESULTS:①CSCs isolated by explants cultures and MACS expressed CD117,which grew in disperse with smaller body.② The expression of cTnT and connexin43 were significantly higher in the co-culture group than that of the control group at days 5 and 7(P

Objective: To optimize the formula and preparation technology of cantharidin chitosan bioadhesive microspheres.Methods: The microspheres were prepared by a spray drying method.A single-factor experiment was used to study the effect of chitosan with different molecular weight on the gastric mucosa adhesion rate were also studied, and the effects of different drug loading ratio, concentration of chitosan acetic acid solution and speed of peristaltic pump on the loading rate were also studied.The cantharidin entrapment efficiency as the studying index, a response surface method was used to further optimize the formula.Results: The best formula and preparation process of cantharidin chitosan bioadhesive microspheres were as follows: the concentration ratio of antharidin to chitosan was 19.83% , the concentration of chitosan acetic acid solution was 0.77% , and the peristaltic pump flow rate was 9.225 ml·min-1.With the best formula, the entrapment efficiency of cantharidin chitosan bioadhesive microspheres was 90.14%.Conclusion: The microsphere preparation by the spray drying method is stable and repeatable.Cantharidin chitosan bioadhesive microspheres have high entrapment efficiency and promising biological adhesion.

Objective To investigate the expressions of NBS1 mRNA and protein in the salivary gland of irradiated rats and explore the role of NBS1 in the repair of radiation injury of salivary gland epithelial cells.Methods Eighty rats were randomly divided into two groups for radiation and control (n =40 each).The rats were fractionally exposed to 3 Gy of 60Co γ-rays once in two days,leading to an accumulation dose of 3,6,9,12,15 Gy.The sham-irradiated controls were anesthetized in parallel but without irradiation.After 2-4 h of irradiation,the rats were sacrificed,IHC and RT-PCR were used to detect the expressions of NBS1 protein and mRNA in parotid and submandibular glands,and the ultra-structural changes in the glands were observed by a transmission electron microscopy.Results After irradiation,the salivary glands became atrophy and the parotid gland cells were damaged more serious than the submandibular gland cells.Compared with the controls,with the groups of dose,at 9,12,15 Gy in parotid gland (t =7.10,17.93,20.86,P ＜ 0.05),at 12,15 Gy in the submandibular gland (t =3.13,7.53,P ＜0.05),the expression of NBS1 mRNA was reduced.With the groups of dose at 9,12,15 Gy in paretid gland (t =4.29,17.91,91.29,P ＜ 0.05 ),the dose at 12,15 Gy in submandibular gland ( t =4.61,11.84,P＜0.05),the expression of NBS1 protein in serous cells,and the dose at 12,15 Gy in parotid gland ductal epithelial cell ( t =3.09,5.62,P ＜ 0.05) were reduced.But in the ductal epithelial cells as well as muoass cells in the submandibualr gland were steadily.Conclusions After irradiation,NBS1 at both protein and mRNA levels was dropped in the salivary gland of rats,which might contribute to the repair of radiation injury of salivary gland.

Objective To evaluate the histologic changes in the dermis and the changes of the rate of type Ⅲ and type Ⅰ collagen by the radiofrequency device. Methods The effects of radiofrequency current on the dermis were observed. Ten rabbits were treated by radiofrequency, and the histologic change in the dermis were observed by H-E staining and Sirius red staining. Results After RF treatment, the fibers in the dermis appeared more compact and the quantity of the type Ⅲ (red) and type Ⅰ (green) collagen were both increased. The fibers in the dermis appeared more compact and the rate of type Ⅲ and type Ⅰ collagen was increased. It was also found that a significant proliferation of dermal collagen was observed in 8 days after treatment. As time went by, the proliferation of dermal collagen was more pronounced, and the rate of type Ⅲ was increased. Conclusion The radiofrequency current can increase the quantity of collagen in the dermis and increase the rate of type Ⅲ and type Ⅰ collagen, which may be one of the key mechanisms of facial rejuvenation by RF.

Objective: To explore the points selection pattern of acupuncture for sleep apnea syndromes by data mining technique. Methods: Clinical literature about acupuncture therapy for sleep apnea syndromes was derived from China National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP), PubMed and Science Direct between the time that databases were created and March 25th,2017. Relevant excel database was established and descriptive studies and association rules were analyzed. Results: The most frequently used point was Lianquan (CV 23) and the most frequently used meridian was the Stomach Meridian. The analysis of association rules showed that the clinical choice of acupuncture points was highly correlated, among which the combination of the highest degree of confidence and the highest degree of support was Shenmen (HT 7) and Sishencong (EX-HN 1); Lieque (LU 7), lianquan (CV 23) and Zhaohai (KI 6). Conclusion: Acupuncture treatment of sleep apnea syndromes has specific selection rules of points, providing certain references for clinical and scientific research.

An actinomycetes which produced soluble blue pigment was isolated from the soil sample in Nanjing,China.Based on its cell chemical characteristics and 16S rDNA sequence we found that its cell wall contained L-diaminopimelic acid and glycine,the whole cell hydrolysates contained glucose and ribose,whole cell contained fatty acid from C14 to C17 with 12-methyltetradecanoic(anteiso-15) and 14-methylpentadentadecanoic acid(iso-16) as the major components.The results shown that,it belongs to the genus Streptomyces.Phylogenetic tree of 16S rDNA sequences indicated that all strains were clustered into 9 branches.All strains that could produce blue pigment were clustered into 2 branches,they were S.coelicolor、S.cyaneus.The isolate closely related to Streptomyces indigocolor with a similarity of 99.4% fell into S.cyaneus branch.

Background Antagonists against vascular endothelial growth factor (VECF) play key roles in treating and preventing neovascular ophthalmopathy. As a novel anti-angiogenic factor, insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) might be an antagonist against VEGF in eye. Objective This study was to explore the inhibitory effect of IGFBP-rP1, a novel anti-angiogenic factor, on VEGF-induced retinal angiogenesis in vitro. Methods The retina-choroid endothelial cell line ( RF/6A ) was cultured in DMEM containing 10% fetal bovine serum. Culture cells were divided into control group(free-serum culture group) ,10mg/L VEGF culture group and different concentrations of IGFBP-rP1 (50,100,200 mg/L) +10 mg/L VEGF group. The expression of IGFBP-rP1 in the cells was detected by immunofluorescence assay. The proliferation and migration of RF/6A cells were evaluated using MTS colorimetric assay and the chemotactic motility assay, respectively. Flow cytometry was used to detect the apoptosis of RF/6A cells. Results The immunofluorescence assay RF/6A cells showed the green fluorescence in cytoplasm and red fluorescence in nuclei after cells were exposed to any concentration of IGFBP-rP1 ,but only red fluorescence was seen in nuclei in control cells. After stimulation of 10 mg/L VEGF,the proliferation value (A490) was elevated and the numbers of cell migration were increased in comparison with control group (t = -15. 191, P = 0. 000; t = -21. 274, P = 0. 000 ) , but the cellular apoptosis rate was lower than the control group (t - 10. 228, P = 0. 000 ) . After treated with various concentrations of IGFBP-rP +10% VEGF, the proliferation and migration of RF/6A cells were significantly decreased in comparison with only 10% VEGF group (F = 534. 158,P = 0. 000;F = 2742. 323,P = 0.000,respectively) ,and the inhibitory effects were gradually enhanced with the increase of IGFBP-rP1 levels (P<0. 05). The apoptosis rate of RF/6A cells in 50,100 and 200 mg/L + 10 mg/L VEGF groups increased by ( 1. 26±0. 04)% ,( 1. 50±0. 07)% and ( 1. 93±0. 27)% respectively,showing significant differences among different groups ( F = 274. 273, P = 0. 000). Conclusion IGFBP-rP1 inhibits the proliferation and activity of retina and choroid endothelial cells induced by VEGF at a concentration-independent manner. It appears to be as a novel endogenous inhibitory factor in retinal angiogenesis.

AIM: To investigate the expression of EMMPRIN, MMP1, MMP9 and TIMP2 in retinoblastoma (RB) and normal retinal tissues and their clinicopathological significance and interrelationship.METHODS: Envision immunohistochemistry stainings of EMMPRIN, MMP1, MMP9 and TIMP2 were performed in 30 enucleated eyeballs with retinoblastoma and 15 specimens of normal retina tissue, which had been routinely imbedded with paraffin.RESULTS: Positive rate of EMMPRIN, MMP1, MMP9 expression was higher in RB tissue than in normal control (P<0.01), while TIMP2 expression was lower in RB than in normal retinal tissue (P<0.01). Samples from RB cases of clinical stage Ⅰ, differentiated type, and life span≥2 years had lower positive rate in expression of EMMPRIN, MMP1, MMP9 than those from RB cases of clinical stage Ⅲ, undifferentiated type, and life span<2 years (P<0.05 or P<0.01), while samples from RB cases of differentiated type, optic nerve unaffected, and life span≥2 years had markedly higher positive rate in expression of TIMP2 than those from RB cases of undifferentiated type, optic nerve involved and life span<2 years (P<0.05 or P<0.01). In RB tissues, EMMPRIN, MMP1, MMP9 expressions were highly consistent (P<0.05), whereas TIMP2 expression is highly inconsistent with EMMPRIN, MMP1, MMP9 expression levels (P<0.05).CONCLUSION: The expression level of EMMPRIN, MMP1, MMP9 and TIMP2 may be an important marker of RB progression, invasion and prognosis. There exist internally mutual regulation relations among them.

ObjectiveTo study a fast and feasible system in clinical application of computer aided socket design and manufacture. MethodsThe biomechanical index were compared of traditional hand-made prosthetic sockets and of computer aided design and manufacture ones based on 3D scanning and reverse engineering. ResultsThe index from 3 cases wearing the computer aided design and manufacture socket prostheses appeared similar or better in static mechanical parameters, walking kinetic parameters and stump-socket interface pressure than they wearing traditional hand-made ones. ConclusionThis computer aided socket design and manufacture system can meet patients' needs in using their prostheses.

This study was purposed to investigate the amplification of CD3(-)CD56(+)NK cells in umbilical cord blood and their change of immunophenotype and cytotoxicity after stimulation with IL-2 and IL-15. Mononuclear cells were isolated from umbilical cord blood and cultured in serum-free medium supplemented with IL-2 or (and) IL-15 for 14 d. The subset level of CD3(-)CD56(+)NK cells and expression of CD16, CD62L, NKG2A, NKG2D, NCR44, NCR46, granzyme B and perforin were analyzed by flow cytometry. The cytotoxicity of NK cells to K562 was detected by WST-1 method. The results showed that NK cells stimulated with IL-2, IL-15 and IL-2/IL-15 were amplified by 10.78 ± 2.51, 10.42 ± 3.72, and 10.54 ± 6.24 times respectively after 14 d, there was no statistically significant difference between these three groups. The expression of CD16 decreased obviously in NK cells after amplification; there was significant difference between IL-2 and IL-15 groups. The expression of CD62L was not changed statistically after stimulation with cytokines, the IL-2 down-regulated the expressions of NKG2A and NCR46, while IL-15 showed the opposite effect. IL-2 or IL-15 displayed upregulation effect on the expression of NKG2D, perforin and NCR44, but there was statistically significant difference between effects of these two cytokines. IL-15 up-regulated the expression of granzyme B on NK cells. The cytotoxicity of NK cells stimulated and amplified by cytokines significantly increased, but there was no statistically significant difference between IL-2 and IL-15. It is concluded that IL-2 or IL-15 can effectively amplify umbilical cord blood NK cells under serum-free conditions. Although the immunophenotype associated with NK cells function showed different characteristics between them, however, cytotoxicity of NK cells increased obviously after amplification and there is no statistically significant difference between effect of these two cytokines, their synergistic effect is not obvious. The cytotoxicity of NK cells is the result from combined effect of all active molecules.
Cells, Cultured ; Cytotoxicity, Immunologic ; drug effects ; Fetal Blood ; cytology ; Flow Cytometry ; Humans ; Immunophenotyping ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; K562 Cells ; Killer Cells, Natural ; drug effects