BACKGROUND:In the cytotoxicity test of the mussel adhesive protein dressing for wound healing, because of positive charge properties of the protein, when extracting ratio is 1:9, the cel s exhibit poly-group phenomenon that results in errors in the cytotoxicity test of mussel adhesive protein samples.
OBJECTIVE:According to the existing standards, to improve the leaching proportion and pretreatment of mussel adhesive protein dressing for wound healing based on the special properties and working condition of mussel adhesive protein.
METHODS:(1) Extract method:Extract solution of mussel adhesive protein dressing was prepared with mussel adhesive protein dressing and cel medium at extracting ratios of 1:9 and 1:131. Then, L929 cel s were cultured in extract solutions of mussel adhesive protein dressing, natural latex and high-density polyethylene, respectively. (2) Method of direct contact:Distil ed water, solutions of mussel adhesive protein dressing, dimethyl sulfoxide and cel medium were used to culture L929 cel s.
RESULTS AND CONCLUSION:When the extracting ratio was 1:9, the cel s agglomerated which is not suitable for cytotoxicity test. When the extracting ratio was 1:131, flocculated sediment and cel aggregation disappeared, the cytotoxicity test results showed no cytotoxicity with higher reliability. Direct contact method showed the samples had no cytotoxicity. The extract method with adjusted extracting radio or direct contact method can be applied to test the in vitro cytotoxicity of mussel adhesive protein dressing for wound healing.

Objective To conduct a descriptive epidemiological study on thyroid cancer incidence in the urbanarea of Tianjin during 2002 - 2006 and to analyze the secular trend of incidences based on previous data of 26 years and those from Shanghai and the United States. Methods Annual percent change (APC%) of thyroid cancer incidence and the rank of all-site cancer profiles were estimated. The secular trend of different histological incidences and the proportion changes were analyzed. Results The incidence of female thyroid cancer was 6. 55/100000 in 2006, ranking the eighth in the female cancer profile. The incidence of thyroid cancer showed an increased secular trend in recent 26 years, especially a 5.7-fold increase in the papillary thyroid cancer (PTC). The incidence for female increased by 267% in 2006 compared to that in 1981, APC% was 3. 1%. The variation ranked second among all the female tumors. The incidences of thyroid cancer in Shanghai and the United States also elevated with time.Conclusions Thyroid cancer has been one of female common tumors from a once less common tumor,especially the proportion of PTC is expanding. It suggests that further study on the risk factors and preventive efforts should be made.

Objective To observe the clinical efficacy ofvitrectomy combined with modified inverted internal limiting membrane (ILM) flap covering technique for complicated macular hole (MH).Methods This is a retrospective case series.Twenty-one eyes of 20 patients who underwent vitrectomy combined with modified inverted ILM flap covering technique were enrolled in this study.Among these eyes,9 eyes were idiopathic MH (IMH),with the mean basal diameter of (1 188.3 ± 155.1) μm,minimum diameter of (626.9± 86.2) μm,logarithm of the minimum angle of resolution (logMAR) best corrected visual acuity (BCVA) of 1.1 ± 0.3;2 eyes were MH with high myopia,with the mean basal diameter of (696.5 ± 232.6) μm,minimum diameter of (259.0±69.3) μm,logMAR BCVA of 1.3;5 eyes were high myopia MH with retinal detachment (RD),with the mean BCVA of 1.5 ± 0.1;3 eyes were rhegmatogenous RD (RRD) with MH,with the mean logMAR BCVA of 1.6;2 eyes were MH after vitrectomy for RRD,with the mean basal diameter of(1 606.0±69.3) μm,minimum diameter of (909.0±387.5) μm,logMAR BCVA of 1.6.All patients received 23G or 25G vitrectomy after removal of posterior vitreous cortex intraoperatively.Indocyanine green staining assisted circle-wise ILM peeling was performed.ILM of diameter 1.5 disc-diameters around fovea was residual and loosened;perfluoronoctane assisted inverting superior or temporal residual ILM covering on macular hole.C3F8,gas or silicone oil tamponade was performed at the end.BCVA and hole closure were followed up for 1-4 months.C3F8,gas or silicone oil was tamponaded at the end.BCVA and hole closure were followed up for 1-4 months.Results MH of 21 eyes were closed after surgery.Nine IMH were closed at type Ⅰ,with U shape closure in 7 eyes,V shape closure in 2 eyes.Two eyes of MH with high myopia,3 eyes of RRD with MH,2 eyes of MH after vitrectomy for RRD were closed at type Ⅰ of U shape.Five eyes of high myopia MHRD including MH closure at type [of U shape 3 eyes,type Ⅱ of W shape 2 eyes.The mean logMAR BCVA of IMH,MH with high myopia,high myopia MHRD,RRD with MH,MH after vitrectomy for RRD eyes were 0.8±0.3,0.9±0.2,1.4±0.1,0.7±0.3,0.9 ± 0.2,respectively.The mean postoperative logMAR BCVA in IMH eyes was improved compared preoperative one (P=0.02).There was no obvious change of pre-and postoperative logMAR BCVA in MH with high myopia,high myopia MHRD,RRD with MH,MH after vitrectomy for RRD eyes (P=0.18,0.10,0.11,0.18).Conclusion Vitrectomy combined with inverted ILM flap covering technique for complicated MH is an effective method to improve the success rate of MH closure and the visual function.

Objective To study the expression of transforming growth factor -β1 (TGF -β1 )in myocardial tissue and the concentration of serum B -type natriuretic peptide (BNP)in myocardial remodeling of SD rats induced by isoproterenol (ISO)and the effects of carvedilol intervention.Methods According to random number table,30 male SD rats were divided into 3 groups:(1 )after 9 g/L saline was injected with 5 mL/(kg·d)for 1 0 days,9 g/L sa-line was used at the dose of 1 0 mL/(kg·d)by way of gavage for 4 weeks in the control group (n =1 0);(2)after ISO was injected with 5 mg/(kg·d)for 1 0 days,9 g/L saline was used at the dose of 1 0 mL/(kg·d)by way of gavage for 4 weeks in the model group (n =1 0);(3)after ISO was injected with 5 mg/(kg·d)for 1 0 days,carvedilol was used at the dose of 1 0 mg/(kg·d)by way of gavage for 4 weeks in the treatment group (n =1 0).Then,all the rats were killed and the cardiac weight index (CWI)was measured.The pathological changes of myocardial tissue were ob-served through HE staining and Masson staining.The mRNA expression of TGF -β1 was detected by adopting reverse transcription -polymerase chain reaction.The protein of TGF -β1 was detected by means of immunohistochemistry and Western blot.The concentration of serum BNP was tested by adopting enzyme -linked immuno sorbent assay.Results (1 )Pathological changes by light microscope:no significant pathological changes were observed in myocardial tissue of the control group.The denaturization,hypertrophy,edema,necrosis of myocardial cells and collagen fibers increased in myocardial tissue of the model group were more serious than those of the treatment group.(2)CWI:CWI of the model group[(3.31 ±0.07)mg/g]was significantly higher than that of the treatment group[(3.03 ±0.04)mg/g],and CWI of the treatment group was significantly higher than that of the control group[(2.98 ±0.1 0)mg/g].There was significant difference among 3 groups (F =54.383,P =0.000).There was also significant difference between any 2 groups (all P <0.01 ).(3)The level of TGF -β1 of myocardial tissue:the level of TGF -β1 of the model group was significantly higher than that of the treatment group,and the level of TGF -β1 of the treatment group was significantly higher than that of the control group by using three detection methods.There was significant difference among 3 groups (F=1 3.62,P =0.000).There was also significant difference between any 2 groups (all P <0.01 ).(4)The concentration of serum BNP:the concentration of serum BNP of the model group[(61 .1 3 ±2.00)ng/L]was significantly higher than that of the treatment group[(57.08 ±1 .52)ng/L],and the concentration of serum BNP of the treatment group was sig-nificantly higher than that of the control group[(51 .56 ±1 .80)ng/L].There was significant difference among 3 groups (F =72.81 ,P =0.000).There also was significant difference between any 2 groups (all P <0.01 ).Conclusions The expression of TGF -β1 in myocardial tissue was up -regulated and the concentration of serum BNP was increased in my-ocardial remodeling of SD rats induced by ISO.The carvedilol intervention can down -regulate the expression of TGF -β1 and decrease the concentration of BNP,then inhibit myocardial remodeling,and improve cardiac function.

OBJECTIVETo evaluate the therapeutic effect of vascular endothelial growth factor (VEGF) and tumor necrosis factor receptor (TNFR) on avascular necrosis of the femoral head in rabbits.

METHODSAvascular necrosis of the femoral head was induced in 26 New Zealand white rabbits by injections of horse serum and prednisolone. The rabbits were then divided into VEGF/TNFR treatment group, VEGF treatment group, and untreated model group, with another 4 normal rabbits as the normal control group. In the two treatment groups, the therapeutic agents were injected percutaneously into the femoral head. Enzyme-linked immunosorbent assay was performed to determine the concentration of TNF-alpha in rabbit serum followed by pathological examination of the changes in the bone tissues, bone marrow hematopoietic tissue and the blood vessels in the femoral head.

RESULTSCompared with the model group, the rabbits with both VEGF and TNFR treatment showed decreased serum concentration of TNF-alpha with obvious new vessel formation, decreased empty bone lacunae in the femoral head and hematopoietic tissue proliferation in the bone marrow cavity.

CONCLUSIONPercutaneous injection of VEGF and TNFR into the femoral head can significantly enhance bone tissue angiogenesis and ameliorate osteonecrosis in rabbits with experimental femoral head necrosis.

Animals ; Drug Therapy, Combination ; Female ; Femur Head Necrosis ; chemically induced ; drug therapy ; Male ; Rabbits ; Random Allocation ; Receptors, Tumor Necrosis Factor ; therapeutic use ; Tumor Necrosis Factor-alpha ; blood ; Vascular Endothelial Growth Factor A ; therapeutic use

The GFP(green fluorescence protein)-streptavidin(SA) bi-functional fusion protein was generated and characterized in order to demonstrate novel platform for efficiently and durably modifying the cell surface with SA-tagged bi-functional proteins.The GFP-SA/pET24d construct was generated and expressed in BL21(DE3) host bacteria at the high level.The recombinant protein GFP-SA was purified through the Ni-NTA affinity chromatography,and then refolded.After biotinylation B16 tumor cells were modified with GFP-SA bi-functional fusion protein and then subjected to fluorescent microscopy and FACS analysis.The effect of surface modification on the viability and growth of B16.F10 tumor cells was evaluated by MTT staining.The GFP-SA recombinant fusion protein was expressed in BL21(DE3) at about 20 % of total bacterial proteins.The GFP-SA bi-functional fusion protein exhibited the bi-functionality,i.e.,SA-mediated high-affinity binding to biotinylated cell surfaces and GFP-emitted green fluorescence.The cell surface modification with GFP-SA bi-functional fusion protein did not affect the viability and growth of the modified B16.F10 tumor cells significantly.The GFP-SA bi-functional fusion protein was obtained and could be displayed efficiently on the surface of the biotinylated B16.F10 tumor cells through the specific and tight interaction between streptavidin and biotin,thus can be used as good trace protein and experimental control in the development of surface-modified tumor vaccine.

Vascular endothelial growth factor 121 (VEGF121) was expressed as inclusion bodies by recombinant Escherichia coli. High concentrations of both biomass (46 g dry cell/L) and VEGF121 inclusion bodies (4.5 g/L) were obtained by applying a high-cell-density culture. After the inclusion bodies were washed and dissolved, VEGF121 was refolded at 0.2 mg/ml by ultrafiltration in refolding buffer with a yield of 81%. Renatured VEGF121 was purified by anion chromatography and Sephacry S-100 chromatography with purity higher than 95% and fnal purification yield of 31%. The purified VEGF121 could stimulate the proliferation of human umbilical vein endothelial cells as demonstrated by a biological activity assay.

Objective To explore the morphological changes of the brain structure in youngsters first-episode depression.Methods Collecting 1 6 youngsters patients with first-episode depression and 1 6 healthy volunteers,separately examining morphological changes in their brain regions by MRI,and then analyzing data by Voxel-Based Morphometry in order to observe the damaged brain areas in depression patients.Results Comparing with control group,gray mater volume of the left dorsolateral prefrontal cortex and right fusiform gyrus decreased in depression patients.The gray matter density of left dorsolateral prefrontal cortex showed negative correlation with HAMD scores.Conclusion The gray matter decreased in left dorsolateral prefrontal cortex and right fusiform gyrus in youngsters first-episode depression patients.In addition,the gray matter density of left dorsolateral prefrontal cortex had negative correlation with HAMD scores.

The objectives of this study are to prepare resveratrol loaded mixed micelles composed of poloxamer 403 and poloxamer 407, and optimize the formulation in order to achieve higher drug solubility and sustained drug release. Firstly, a thin-film hydration method was utilized to prepare the micelles. By using drug-loading, encapsulation yield and particle size of the micelles as criteria, influence of three variables, namely poloxamer 407 mass fraction, amount of water and feeding of resveratrol, on the quality of the micelles was optimized with a central composite design method. Steady fluorescence measurement was carried out to evaluate the critical micelle concentration of the carriers. Micelle stability upon dilution with simulated gastric fluid and simulated intestinal fluid was investigated. The in vitro release of resveratrol from the mixed micelles was monitored by dialysis method. It was observed that the particle size of the optimized micelle formulation was 24 nm, with drug-loading 11.78%, and encapsulation yield 82.51%. The mixed micelles increased the solubility of resveratrol for about 197 times. Moreover, the mixed micelles had a low critical micelle concentration of 0.05 mg · mL(-1) in water and no apparent changes in particle size and drug content were observed upon micelles dilution, indicating improved kinetic stability. Resveratrol was released from the micelles in a controlled manner for over 20 h, and the release process can be well described by Higuchi equation. Therefore, resveratrol-loaded poloxamer 403/407 mixed micelles could improve the solubility of resveratrol significantly and sustained drug release behavior can be achieved.
Drug Carriers ; chemistry ; Fluorescence ; Kinetics ; Micelles ; Particle Size ; Poloxamer ; chemistry ; Solubility ; Stilbenes ; chemistry ; Water

Objective To detect the expression of inducible nitric oxide synthase (iNOS) and heme oxygenase-1(HO-1) in human colon of irritable bowel syndrome(IBS), aim at finding out the role of gas messenger molecule such as nitric oxide(NO) and carbon monoxide(CO) in the pathogenesis of IBS. Methods The expression of iNOS and HO-1 in human colonic tissue specimen of enteroscope was detected by En vision and SABC immunohistochemistry each in 34 cases of IBS and 16 cases of normal control group. Results The expression of iNOS and HO-1 expressed mainly in the colonic mucous membrane epithelia, and the expression in IBS group was higher than that in normal control group(P