Objective:The study aims to screen chemosensitivity-associated proteins in colorectal carcinoma tissues by using two-dimensional gel electrophoresis (2-DE) and mass spectrometry,then identify some differentially-expressed proteins. Methods:The patients with advanced colorectal carcinoma were confirmed by clinical diagnosis. Fresh carcinoma specimens were collected by biopsy and preserved in liquid N2. The tissues were classified into two groups: high sensitivity group (HS) and low sensitivity group (LS) based on drug sensitivity test. The total proteins were extracted and separated by 2-DE. The images were composed, compared, and differentially analyzed to identify the proteins with differential expression in HS and LS groups. Then the differentially-expressed protein spots were incised from the gels and digested by trypsin. The peptide mass fingerprintings (PMF) was acquired after matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and the proteins were identified by data searching in the Mascot database. Two proteins with differential expression were detected by Western blotting.Results:The 2-DE spectrum of HS and LS groups were established. Most protein spots were distributed in the area with pH 4-8 and relative molecular weight of (20-100)×10~3. The average number of the protein spots was 842±23 in HS group and 793±19 in LS group,respectively. The mean matching rate was 90.7%. The number of differentially-expressed dots between HS and LS group was 79.00±13.56. Thirty protein dots were selected for mass spectrum and bioinformatic analysis, and 9 proteins were identified. Conclusion:Colorectal carcinoma with different chemosensitivity had differential protein expression profiles. The differentially expressed proteins may be associated with chemosensitivity and could be used for prediction of chemosensitivity of colorectal carcinoma.

This paper focuses mainly on the study of optimal fermentation conditions of S-adenosyl-L-methionine.Effects of carbon sources,nitrogen sources,inorganic constituents,growth factors and adding time of L-methionine on the yield,the content and biomass of S-adenosyl-L-methionine are studied.And ingredients of the culture medium are also optimized by the method of uniform design.The final optimum culture medium contains: glucose 30 g,Yeast powder 11 g,(NH_4)_2SO_4 12 g,K_2HPO_4?3H_2O 5 g,KH_2PO_4 10 g,MnSO_4?H_2O 0.09 g,ZnSO_4?7H_2O 0.14 g,MgCl_20.5 g,CaCl_2 0.3 g,CuSO_4 0.005 g per liter. Using that optimum culture medium,the yield of S-adenosyl-L-methionine can reach 0.9 g/L in Erlenmeyer flask which is 30 % higher than before.Experiment on 5 L fermenter reveals that the accumulation of S-adenosyl-L-methionine can reach 2.66 g/L.Biomass is 23.4 g/L.

Objective To report the outcome of emergency repair traumatized limbs by vascularized free tissue transplantation.Methods From April 1988 to August 2004,86 patients,58 men and 28 women,had undergone emergency vascularized free tissue transplantation to have their injured limbs repaired in 54 cases and the missing thumbs reconstructed in 32.The patients aged from 5 to 55 (mean 27.9) years. The transplants included latissimus dorsi myocutaneons flap,anterolateral femoral skin flap,medial crural skin flaps,dorsal pedal flaps,medial plantar flap,composite tissue mass of the discarded limbs and big toe skin- nail flap.The operations were performed 1 to 5 days after injuries.Results Postoperative vascular crises occurred in 8 cases and were all followed by exploration with successes in 5 cases while failure in 3.The total survival rate of the transplants was 96.5% (83/86).In this series all the patients were followed up for 1 to 16 years with a mean of 7.5 years only to reveal satisfying functional recovery in all the repaired limbs and an ex- cellent and good rate of 87.5% in the reconstructed thumbs.Conclusion Emergency vascularized free tis- sue transplantation is an effective way to repair a traumatized limb and to reconstruct a traumatically missing thumb.

AlM: To find out the weaknesses of the cultivation of the Chinese ophthalmology physicians and the gap between Chinese and the international ophthalmology physicians, so that provide the advice on the future cultivation of the Chinese ophthalmology physicians.
METHODS: The passrate of the 2013 lCO examinations taken by worldwide examiners by common statistical methods was analyzed.
RESULTS:The results indicated that the test scores of Chinese candidates' were lower than that of the international average level, there was a obvious gap existed between Chinese and other countries' ophthalmology physicians. lt showed that Chinese candidates were not quite adaptable to this examination, basic science and clinical level needed to be improved.
CONCLUSlON:lt may shows that the effects on the mid-anaphase of our country's ophthalmology residency training are not so good, which area we should pay more attentions.

Esophageal and Gastric Varices ; etiology ; therapy ; Hemodynamics ; Humans ; Hypertension, Portal ; complications ; therapy ; Liver Cirrhosis ; complications ; therapy ; Portal Vein ; physiopathology

The study was aimed to investigate the possible effects of 8-chloroadenosine 3', 5'-monophosphate (8-Cl-cAMP) on the multiple myeloma cells. The multiple myeloma cell line RPMI8226 was used as in vitro models. The effect on growth inhibition of RPMI8226 cells was evaluated by cell growth and viability curve. DNA fragment was visualized by agarose gel electrophoresis. The amount of apoptosis cells was measured by flow cytometry. Meanwhile Western blot assay were used to detect the change of several key cell cycle regulatory proteins CDK2 and cyclin E in these cells before and after the treatment. The results showed that low dose 8-Cl-cAMP (1-30 micromol/L) inhibited the proliferation and viability of RPMI8226 cells significantly. Agarose gel electrophoresis of DNA revealed the apoptosis characteristic "ladder" pattern. Apoptosis was also confirmed by flow cytometry. In addition, 8-Cl-cAMP was able to inhibit the cell growth through modulating expression of cell cycle regulators CDK2 and cyclin E. It is concluded that 8-cl-cAMP inhibits the proliferation and induce apoptosis of multiple myeloma cells effectively.
8-Bromo Cyclic Adenosine Monophosphate ; analogs & derivatives ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase 2 ; metabolism ; Humans ; Multiple Myeloma ; pathology

The yield of S-adenosyl-L-methionine (SAM) by saccharomyces cerevisiae fermentation was affected by the strategy of feeding L-methionine. The effects that feeding strategies and the amount of precursor L-methionine had on the production of SAM by saccharomyces cerevisiae G14 were investigated. The results showed that feeding L-methionine could obviously improve the accumulation of SAM, and both the biomass and SAM yield relied heavily on different feeding strategies. In our work, it was found that total amount of L-methionine added should be no less than 0.7g per 10 grams of dry cell weight. Five different feeding strategies had been investigated in our experiment, and such comparison indicated that favorable results could be achieved as the biomass reached the status of high cell density (120g/L). If 9 grams of the precursor L-methionine was introduced once and for all, the accumulation of SAM reached maximum of 4.31g/L at the 18th hour after addition; if the precursor amino acid was fed at a rate of 2g/h in 5 h, maximum yield of 4.98g/L was achieved at the 28th hour after feeding. Thus high cell density fermentation can be successfully applied to SAM production by Saccharomyces cerevisiae with the consequence of over 130g/L of biomass gained using the above two strategies.
Bioreactors ; microbiology ; Cell Culture Techniques ; methods ; Culture Media ; Fermentation ; Methionine ; metabolism ; S-Adenosylmethionine ; biosynthesis ; Saccharomyces cerevisiae ; growth & development ; metabolism

Objective To explore the effects of amniotic lacrimal duct stenting on the prevention of dry eye in castrated rabbits.Methods Thirtysix healthy male rabbits were selected,the third eyelid were cut off and antiinfection treatment were given,which were randomly divided into 3 groups (12 cases in each group),the castrated male rabbits models were made.Among them,group A was negative control group,group B was dry eye model group,group C was group of lacrimal amniotic membrane group.At 2 weeks before implantation of amniotic lacrimal duct stent,2 weeks,4 weeks and 6 weeks after implantation,the fluorescent (FL) examination,Western blot,Schirmer I examination,immunofluorescence staining and corneal confocal microscopy were performed.Results The levels of tear secretion and FL in the three groups among different time points were significantly different (F=7.126,P =0.009;F =9.658,P =0.016),and there were significant differences among three groups (F =12.582,P =0.005;F =13.187,P =0.013).The tendency of tear secretion and FL in the three groups were also significantly changed (F =8.531,P =0.007;F =10.652,P =0.019).The epithelial basal cells at 6 weeks after implantation in three groups were 3811 ±414,3820 ± 314,2789 ± 353,and the density of inflammatory cells was 266 ±28,266 ± 29,67 ± 13,there were significant differences among three groups (F =13.442,P =0.012;F =9.231,P =0.021).The K1 6 staining in the duct epithelium were negative,and the expression of α-SMA in the lacrimal duct tissue of group A,B and C was not changed at all time points after implantation of amniotic lacrimal stent,and there was no significant difference (F =14.681,P =0.002).Conclusion The amniotic lacrimal stent implantation has certain effect on the prevention of dry eye in rabbit.

This study was aimed to investigate the possible effects of cyclic adenosine monophosphate (cAMP) analogue 8-(4-chlorophenylthio) adenosine 3', 5'-cyclic monophosphate (8-CPT-cAMP) on the M(2b) subtype of acute myeloid leukemia (AML-M(2b)) cells. AML-M(2b) is characterized by the non-random chromosome translocation t (8; 21) (q22; q22), through which AML1 (acute myeloid leukemia 1) gene on chromosome 21 is fused with ETO (eight twenty-one) gene on chromosome 8, coding correspondent AML1-ETO fusion protein, which plays a crucial role in the leukemogenesis of AML-M(2b). The AML-M(2b) cell line Kasumi-1 cells were used as an in vitro model. The influences of 8-CPT-cAMP on the proliferation and differentiation of Kasumi-1 cells were evaluated according to cellular morphology, changes in cell surface antigen and cell cycle, as well as nitroblue-tetrazolium (NBT) assay. Meanwhile, semi-quantity RT-PCR and Western blot assay were used to detect the degradation of AML1-ETO fusion protein in Kasumi-1 cells before and after the treatment. The results showed that 8-CPT-cAMP (200 micromol/L) could significantly inhibit cell growth and induce differentiation of Kasumi-1 cells. However, it must be pointed out that 8-CPT-cAMP-induced differentiation in Kasumi-1 is not a typical terminal differentiation. Furthermore, 8-CPT-cAMP exerted little influence on the expression of AML1-ETO fusion gene and its product in Kasumi-1 cells. In conclusion, the 8-CPT-cAMP induced differentiation in Kasumi-1 cells. This results may provide experimental and theoretical basis for the breakthrough of differentiation-induced therapy extended to another leukemia.
Cell Transformation, Neoplastic ; drug effects ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; Cyclic AMP ; analogs & derivatives ; pharmacology ; Humans ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; pathology ; Oncogene Proteins, Fusion ; genetics ; metabolism ; RUNX1 Translocation Partner 1 Protein ; Thionucleotides ; pharmacology ; Tumor Cells, Cultured

Adult ; Female ; Hand ; surgery ; Humans ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Toes ; transplantation