BACKGROUND: Fibronectin serves not only as the supporter for cells,but also as an important intracellular linkage, possessing opsonic-like functions. It can promote the phagocytic function of mononucleophages and the repair in inflammation and trauma.OBJECTIVE: Type O plasma was freshly obtained from healthy males in search of the optimal preparation method for lower sampling, convenient and higher-yielding of fibronectin.DESIGN: Open experiment.SETTING: Institute of Traumatic Surgery, Fourth Affiliated Hospital of Guangzhou Red Cross Hospital, Jinan University.MATERIALS: This experiment was carried out in the Institute of Traumatic Surgery of Guangzhou Red Cross Hospital between July 2000 and July 2002. The major materials consisted of type O fresh plasma from healthy males, gelatin, sepharose 4B activated with cyanogen bromide,sephadex G-25, urea, and trishydroxymethylaminomethane.METHODS: Affinity column constituted by gelatin coupling with sepharose 4B activated with cyanogen bromide was used to purify fibronectin. ① Human plasma was added: 150 mL type O plasma was freshly obtained from healthy males and added into the column once by 25 mLat an interval of 20 minutes, the speed of flow was 3.5 mL/min. ② Removing mixed protein: the column was rinsed by Tris-sodium citrate equilibrium liquid (pH 7.5) until the absorbency of flow-out fluid decreased to ＜ 0.02 at 280 nm. Again lmol/L NaCl containing Tris-sodium citrate was used to wash off other mixed proteins. ③ Collecting fibronectin: the column was rinsed by urea-Trispurge Fluid (3 mol/L) to collect fibronectin.④ Removing urea from fibronectin collection: fibronectin collection liquid was filtrated by Sephade G-25 to remove urea. ⑤Disinfection: 0.22 μmgermtight filter was used for disinfection.MAIN OUTCOME MEASURES: ①Results of sodium dodecyl benzene sulfonate-polyacrylamide gel electrophoresis. ② The influence of retention time on the yield of purified fibronectin. ③ The influence of plasma quantity on the yield of purified fibronectin.RESULTS: ① Results of sodium dodecyl benzene sulfonate-polyacrylamide gel electrophoresis: The density of separation gel and condensed gel was 7% and 3%, respectively; fibronectin was electrophoresized into single protein lane. ② The influence of retention time on the yield of purified fibronectin: The yield of fibronectin was (65.24±3.45) %, (74.77±4.05) %,(86.99±4.10) %, and (80.47±3.75) %, respectively, when the loading amount of fibronectin was 150 mL with non-retention time and retention time of 10, 20 and 30 minutes. ③ The influence of plasma quantity on the yield of purified fibronectin: The yield of fibronectin was (72.56±3.63) %,(77.61±3.14) %, (86.99±4.12) %, and (74.67±3.05) % when the column retention time was controlled at minute 20 with the loading amount of 100,130, 150 and 180 mL, respectively.CONCLUSION: In a given column volume of gelatin, the quantity of purified fibronectin was closely related with the plasma retention time in column and the total loading amount of plasma. As a result, the optimal loading amount of plasma was 150 mL, and the retention time was 20 minutes.The preparation method, herein, has been proved to require small amounts of plasma and yield large amounts of fibronectin.

Objective To investigate the effect and mechanism of melatonin on bioactivity of fibroblasts from human hypertrophic scar.Methods Fibroblasts from hypertrophic scar were cultured and incubated with melatonin,melatonin and Luzindole,and Luzindole,respectively,for 24 h,and the media as control.XTT/PMS assay was used to measure the proliferation of fibroblasts,ELISA assay to detect the TGF-β1 production of fibroblasts,and the expression of cell a-SMA,collagen Ⅰ,collagen Ⅲ mRNA were determined with real-time PCR method.Results Compared with the control,melatonin at the concentration of 10-5mmol/L,10-3mmol/L,and 1 mmol/L could inhibit the proliferation of fibroblasts in a does-dependent manner (P＜0.05); melatonin at the concentration of 10-3 mmol/L could significantly decrease the TGF-β1 production and expression of a-SMA mRNA and collagen Ⅰ mRNA in fibroblasts from human hypertrophic scar (P＜0.05) ; the effect of melatonin on fibroblast was significantly blocked by Luzindole (P＜0.05),but melatonin could not inhibit collagen Ⅲ mRNA expression (P＞0.05).Conclusions Melatonin can significantly regulate the biological activity of fibroblasts from human hypertrophic scar through a receptor pathway.

Objective To evaluate the feasibility,safety and effectiveness of modified associating liver partition and portal vein ligation for staged hepatectomy (ALPPS).Methods The published literatures associated with modified ALPPS were pooled from Embase,Pubmed,Medline,Google Scholar databases.The studies were included or excluded depends on our predetermined criteria.We selected data and performd descriptive analysis from the included studies.Results Five articles were included and reviewed.A total of 62 patients underwent five modified procedures,including monosegment ALPPS (m-ALPPS),anterior approach ALPPS,partial-ALPPS,radiofrequency-assisted liver partition with portal vein ligation (RALPP) and associating liver tourniquet and portal ligation for staged hepatectomy (ALTPS).There were 50 (80.6％) patients diagnosed liver metastatic colorectal cancer.The average operation interval of modified ALPPS was between 8 ～ 22 days and growth rate of future liver remnant (FLR) ranged from 48.7％ to 62.3％,the feasibility to perform ALPPS stage 2 was 98.4％.The incidence of severe postoperative complications were between 11.8％ ～33.3％.The 90-day mortality for monosegment ALPPS,partial-ALPPS and RALPP was 0,while the figure was 8.3％ in ALTPS.The in-hospital morbidities were 5.9％ and 8.3％ for anterior approach ALPPS and ALTPS,respectively,which were 0 in the other three modified groups.Clinical response evaluation,including R0 resection rate,overall survival rate,disease-free and recurrence rates were merely presented 83.3％,80％,50％,50％ in m-ALPPS group,while 100％,100％,95％,5％ in modified ALTPS group.Conclusion Modified ALPPS with improved safety is feasible in clinical practice.However,the effectiveness still needs further studies.

BACKGROUND: Simple polyvinyl alcohol (PVA) has limited ability to cell adhesion. There are not generally accepted studies on improved effects of collagen protein modified polyvinyl alcohol on cell adhesion and proliferation.OBJECTIVE: To investigate the feasibility of PVA/type Ⅰ college (COL-Ⅰ) as anterior cruciate ligament (ACL) scaffolds in tissue engineering.DESIGN, TIME AND SETTING: The controlled observation experiment was performed at the Fourth Affiliated Hospital, Medical College. Ji'nan University, Guangzhou Red Cross Hospital, Guangzhou Institute of Trauma Surgery from August 2006 to October 2007.MATERIALS: COL-Ⅰ gel was produced by Guangzhou Institute of Trauma Surgery.METHODS: PVA filature was used to weave fascicular scaffolds. NIH-3T3 cell line and human ACL cells were in vitro incubated, amplified, and then implanted on the PVA/COL scaffolds.MAIN OUTCOME MEASURES: The growth of NIH-3T3 cell line and human ACL cells on the PVA/COL scaffolds and the secretion of extracellular matrix were observed using scanning electron microscope. Cell compatibility of PVA/COL scaffolds was assessed. Mechanics characteristic of PVA/COL scaffolds was measured by using the electric. tensile force apparatus. Mechanical property of PVA/COL scaffolds was analyzed using the SPSS 11.5 software package.RESULTS: NIH-3T3 cell line and human ACL cells on the PVA/COL scaffolds adhered, proliferated, and secreted extracellular matrix. NIH-3T3 cell line highly grew compared with human ACL cells on the PVA/COL scaffolds. The adhered number of NIH-3T3 cell line and human ACL cells was significantly increased on the PVA/COL scaffolds. NIH-3T3 cell line and human ACL cells presented well morphology on the PVA/COL scaffolds. COL-Ⅰ could promote the secretion of extracellular matrix from NIH-3T3 cells, but its effects on human ACL cells were not significant. Tensile force test showed that load-extension curve of the materials was identical to ACL of human and rabbits, and the scaffolds possessed strong flexibility. The maximal load, ultimate stress and elastic modulus were respectively 52.61 N, 14.96 MPa and 202.08 MPa.CONCLUSION: COL-Ⅰ accelerates the adhesion and proliferation of NIH-3T3 cell line and human ACL cells on the surface and in the pore of the PVA/COL scaffolds, promotes the secretion of extracellular matrix from NIH-3T3, and PVA filature material has mechanical property and good cell compatibility.

Objective To investigate the effect of 15d-PGJ2 on the expression of collagen type,CTGF and a-SMA in the hypertrophic scar in the rabbit ear,and the possibility of hypertrophic scar treated by 15d-PGJ2.Methods 18 New Zealand white rabbits were used to establish a hypertrophic scar model on the rabbit ear.The wounds were established as follows:2 cm × 3 cm wounds with total skin loses on the ventral side,2 wounds for each ear,totally 72 wounds.The wounds were randomly divided into the 15d-PGJ2 treatment group and NS control group.20μl 15d-PGJ2 or NS was injected into the ear scar once a day for 7 days.At 7,14 and 21 days after the injection,12 scars of each group were harvested.The expression of collagen type Ⅰ,CTGF and a-SMA was detected by immunohistochemical method.Results Excessive dermal scars on rabbit ears that were similar to human hypertrophic scar appeared in the two groups.Compared with the NS-treated scars group,the 15d-PGJ2-treated scars appeared to be smaller,softer,flatter and lighter in color.The expression of collagen type Ⅰ,CTGF and a-SMA in the 15d-PGJ2 group was significantly decreased as compared with that in the control group at different time points(P＜0.05).Conclusion 15d-PGJ2,the ligand of PPAR-r,can reduce the expression of collagen type Ⅰ,CTGF and a-SMA of hypertrophic scar in the rabbit ears and plays an important role in the prevention and treatment of hypertrophic scar.It may provide a new approach for the treatment of hypertrophic scar in clinical setting.

BACKGROUND: Fibronectin(FN) plays the role of repair in the inflammation. There is no confirmed conclusion whether it can be applied to the refractory corneal epithelial defect.OBJECTIVE: To observe the repair effect of the human plasma FN for the refractory corneal epithelial defect.DESIGN: A controlled experimental study.SETTING: Guangzhou Red Cross Hospital, Guangzhou First People's Hospital, Guangzhou Children's Hospital, Guangzhou Hospital of Traditional Chinese Medicine and Guangzhou Second People's Hos pital.PARTICIPANTS: Totally 383 eyes with the refractory corneal epithelial defect were chosen, of which 309 were in the therapy group, and 74 in the control group.METHODS: The therapy group: Human plasma FN was administered by dropping it into the eyes once every two hours. The controlgroup: 10 g/L celacol M was administered by its dribbling into the eyes once every two hours. Weilesheng was taken orally in both groups, two pills once, three times per day. According to the state of illness, both groups received anti-bacterial or anti-viral treatment and reexamination was given every day or every other day after administration. 10 g/L fluorescein sodium was used to observe the changes of cornea.MAIN OUTCOSE MEASURES: The symptoms, results of staining using fluorescein sodium as well as the corneal epithelial healing of both groups.RESULTS: The symptoms, the results from staining using the fluorescein sodium and the corneal epithelial healing were used for the evaluation. In the therapy group, 309 eyes were followed up and the cure rate was 69.9%. The average therapeutic period was 6.5 days, while those of the control group were 58.1% and 8.7 days respectively. The difference in the curative effect between the two groups was significantly different( P＜0.01 ).CONCLUSION: The application of FN for the refractory corneal epithelial defect displays a more significant effect than conventional treatment.

The preparation of collagen sponges was studied in order to develop tissue engineering scaffolds. Collagen solutions with varying concentrations were obtained by condensing the initial collagen with polyethylene glycol (PEG) at 4 degrees C for different periods of time, and then were freeze-dried to make collagen scaffolds. The porous characteristics of the prepared scaffolds were characterized by use of different methods, including laser scanning confocal microscopy (LSCM), scanning electron microscopy (SEM) and tensile tests. All collagen sponges were shown to have similar interconnected porous structures but were found to have different pore size, porosity, water capacity and the mechanical property, depending on the concentration of collagen solutions. These findings indicate that the way of controlling the concentration of collagen solutions with PEG permits the freeze-drying fabrication of collagen sponges with varying porous features suitable for different tissue engineering purposes.
Collagen ; chemistry ; ultrastructure ; Freeze Drying ; Polyethylene Glycols ; chemistry ; Porosity ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo investigate the protective effect of mannitol therapy on the vital organs and explore the underlying mechanisms in New Zealand rabbits with severe burn injury.

METHODSTwelve New Zealand rabbits with severe burn injury (30% of TBSA) were randomized to receive fluid resuscitation with saline (control) or mannitol therapy starting at 1 h after the injury. Serum and urine samples were collected before and at 1, 4, 8, 24, and 48 h after the injury for detection of TNF-α, IL-6, ALT, AST, GGT, CK, CK-MB, BUN and Cr levels using sandwich ELISA.

RESULTSOne hour after sever burn injury, the serum levels of TNF-α and IL-6 began to increase along with ALT, AST, GGT, CK, CK-MB, BUN and Cr levels. Compared with control group, the rabbits in mannitol group showed significantly higher 48 h urine excretion of TNF-α (145 ± 8 vs 78 ± 1 0 pg/ml, P<0.05) and IL-6 (93 ± 6 vs 40 ± 8 pg/ml, P<0.05) but with lowered serum levels of TNF-α (0.62 ± 0.02 vs 0.83 ± 0.02 pg/ml, P<0.05) and IL-6 (0.45 ± 0.03 vs 0.56 ± 0.03 pg/ml, P<0.05) as well as lowered serum ALT, AST, GGT, CK, CK-MB, BUN and Cr levels (P<0.05).

CONCLUSIONIn rabbits with severe burn injury, mannitol therapy can decrease serum TNF-α and IL-6 levels early after the injury to ameliorate potential functional impairment of the heart, liver and kidneys.

Animals ; Burns ; blood ; drug therapy ; Fluid Therapy ; Interleukin-6 ; blood ; Male ; Mannitol ; therapeutic use ; Rabbits ; Tumor Necrosis Factor-alpha ; blood

In search of the optimal preparation method for large-scale purification of human plasma fibronectin, we adopted affinity chromatography with gelatin and the Sepharose 4B activated with cyanogen bromide to purify fibronectin from type "C" plasma of healthy males, and scanned the best method under the conditions of different amount of plasma loading and different residence time in column. In a given column volume of gelatin, the absorbent was related with the plasma residence time in column and the total amount of plasma loaded. As a result, the optimal loading amount of plasma is 150 ml, and the residence time is 20 minutes. The preparation method, herein, has been proved to require small amount of plasma and yield large amount of fibronectin.
Chromatography, Affinity ; methods ; Cyanogen Bromide ; chemistry ; Fibronectins ; blood ; isolation & purification ; Gelatin ; Humans ; Male ; Sepharose ; chemistry ; Wound Healing