BACKGROUND: Study on bone tissue-engineered material is one of the most successful fields in tissue engineering, but the mechanism on synthesis of artificial bone has not been known in many aspects.OBJECTIVE: To explore the mechanism of collagen and calcium phosphate (CP) in artificial bone synthesis.DESIGN: Single sample experiment was designed.SETTING: Material Research Room of Honghe University.MATERIALS: The experiment was performed in Material Research Room of Honghe University from July to August 2003. The materials included collagen (10 g/L acetic acid solution), calcium chloride, sodium dihydrogen phosphate (SDP), sodium hydroxide (NaOH), Tris, hydrochloric acid and deionized water (DI water).METHODS: Liquid nitrogen freezing and freeze-drying were used to prepare collagen-CP complexes A and B and the samples at different times during mineralization. UV spectrophotometer was used to determine the biomineralized dynamic curve of collagen-CP. Based on law of curve, the different times of sample collection were determined in preparation of electronic microscopic samples. According to electronic microscopic pictures and spectral data, mechanism analysis was carried on.MAIN OUTCOME MEASURES: Morphology of collagen-CP complex and law of its structure with time changeRESULTS: ①Under agitation, collagen-CP complex A was sheaf-like or needle-like in structure manufactured with retarded neutralization. ②Under static state, with biomineralization, collagen-CP complex B was in layered structure at initial phase of mineralization, which was similar to the self-assembled structure of pure collagen and the molarratio of C, O, P and Ca was 7.26: 20: 0: 2. At the end of mineralization, the structure was strip-like in high density with a certain grains and very fine rills and the molar ratio of C, O, P and Ca was 11.02: 22.5:1.06: 2.CONCLUSION: At the early phase of biomineralization, collagen iscoordinated initially with calcium ion, calcium-carrier layered collagen template is formed with the self-assembling of collagen, and then phosphates is combined with calcium ion to manufacture calcium phosphate in the formed template. By controlling agitation and acting time, collagen complex material of reticular and spinal structure is obtained.

The culture medium and cultural conditions of solid-state fermentation of Streptomyces Menmyco-93-63 were tested in this study. The suitable medium which contains rice, sorghum, millet bran, and rice hull with the proportion of 2:2:3:3 was developed for the spore production of Streptomyces Men-myco-93-63 using single substrate screening, mixture substrate screening and orthogonal experiments, and the sporulation was up to 2.52?109 CFU/g. And then, initial charge, initial ratio of water to solid, inoculating quantity, and culture temperature impact to sporulation of Streptomyces Men-myco-93-63 were tested. The favorite cultural conditions are developed as the following: the initial charge is 15 g in 500 mL Erlenmeyer flask; initial ratio of water to solid is 1.7:1.0 (V/W, rice hull excluding), inoculating quantity is 7 mL, culture temperature is 28℃.

Objective To investigate the correlation between specific features on contrast-enhanced CT and its clinical findings of adrenal tuberculosis so as to improve the diagnostic accuracy of the disease. Methods Contrast-enhanced CT features in 30 patients with documented adrenal tuberculosis were retrospectively evaluated blindly for the features of location,size,morphology,attenuation and enhancement patterns on CT images,and compared with clinical and pathological materials.Results The common CT manifestations were as follows:enlargement of the adrenal glands in all 30 cases(bilateral 90%,mfilateral 10%)including mass-like enlargement in 13 cases and enlargement but the contours preserved in 17 cases, heterogeneity(28 cases,93.3%),calcification(17 cases,56.7%),and low attenuation in the center with peripheral enhancement(16 cases,53.3%)of the lesions.After antituberculosis chemotherapy, 5 cases of enlarged adrenal glands decreased in size or returned to normal size and configuration,with disappearance of the central low attenuation and new appearance of dot-like calcification in 2 cases.Cochran Armitage trend test showed there was an increasing tendency of calcification rate with clinical duration(X~2= 7.47,P

OBJECTIVETo observe the effect of umbilical sticking therapy (UST) with Qitou Xiaugu Plaster (QXP) on hemodynamics of portal system in patients with liver cirrhosis.

METHODSOne hundred and twenty patients of liver cirrhosis with portal hypertension were assigned to two groups. On the basis of conventional therapy, UST was applied in the 66 patients in treated group, which was exchanged once every 3 days with an interval of 1-day rest. The 54 patients in the control group were orally administered with propanolol. The therapeutic course for both groups was 1 month. Before and after treatment, the hemodynamic changes in portal or splenic veins were observed by color Doppler ultrasonograph, and the changes of liver function, blood coagulation and patients' subjective symptoms were observed as well.

RESULTSAfter treatment, portal vein diameter and splenic vein diameter significantly decreased (P < 0.05, portal venous flow velocity and splenic venous flow velocity apparently increased (P < 0.05), and portal venous flow apparently decreased in both groups (P < 0.05), while no significant change was found in the splenic venous flow (P > 0.05). The liver function and blood coagulation indexes in both groups were improved. The improvement of clinical symptoms in the treated group was superior to that in the control group.

CONCLUSIONUST with QXP could decrease the portal vein pressure in a short time, with the therapeutic effect comparable to propanolol, and with no adverse reaction.

Adult ; Aged ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hemodynamics ; drug effects ; Humans ; Hypertension, Portal ; drug therapy ; physiopathology ; Liver Cirrhosis ; drug therapy ; physiopathology ; Male ; Middle Aged ; Portal Pressure ; drug effects ; Portal Vein ; drug effects ; physiopathology ; Splenic Vein ; drug effects ; physiopathology ; Umbilicus ; blood supply ; Young Adult

This study was designed to investigate the correlation between autophagy and polarization of macrophages in atherosclerosis (AS) plaque in arteriosclerosis obliterans amputees. Femoral artery specimens from arteriosclerosis obliterans amputees were performed hematoxylin and eosin (HE) staining, oil red O and immunofluorescence staining to observe the morphology of atherosclerotic plaque, phenotype of macrophages and autophagy in plaque; using real-time quantitative RT-PCR technology to detect the mRNA level of M1 and M2 type markers in arterial tissue; to analyze polarized signal pathway and autophagy protein levels in macrophages by Western blotting. Arterial specimens staining showed obvious lipid deposition and obvious infiltration of amount of foam cells and inflammatory cells. Macrophages were mainly expression M1 type in percentage in fibrous plaque. Although both M1 and M2 macrophages were upregulated in atheromatous plaque, the increase was dominant in M2 type in percentage. The level of autophagy was significantly higher in the atheromatous plaque than that of fibrous plaque. The expression of tumor necrosis factor- α (TNF-α), monocyte chemotactic protein-1 (MCP-1), inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) and interleukin-12 (IL-12) mRNA was significantly higher in fibrous plaque than that of atheromatous plaque (P < 0.01 or 0.05), and arginase-1 (Arg-1), transforming growth factor-β (TGF-β), CD163 and interleukin-10 (IL-10) mRNA was significantly lower than that in atheromatous plaque (P < 0.01). The levels of p-STAT1 and NF-κB were significantly increased in fibrous plaque (P < 0.01), while p-STAT6 expression was significantly increased in atheromatous plaque (P < 0.01). The level of LC3-II was significantly higher in atheromatous plaque than that in fibrous plaque (P < 0.01). Macrophages in early atherosclerotic plaque were induced to M1 type through p-STAT1/NF-κB pathway and expressed moderate levels of autophagy; while macrophages in advanced plaques were induced to polarization of M2 type through p-STAT6 pathway. M2 macrophages expressed a higher level of autophagy than M1 macrophages.
Amputees ; Arginase ; metabolism ; Arteriosclerosis Obliterans ; pathology ; Atherosclerosis ; pathology ; Autophagy ; Cell Polarity ; Chemokine CCL2 ; metabolism ; Foam Cells ; cytology ; Humans ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-6 ; metabolism ; Macrophages ; cytology ; NF-kappa B ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Phenotype ; STAT6 Transcription Factor ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Up-Regulation

Objective To explore the expression of silencer of death domains(SODD) and its clinical significance and relationship with phospho-NF-?B-p65 proteins in bone marrow cells of acute lymphoblastic leukemia(ALL)in children,and the expression of SODD and phospho-NF-?B-p65 in Jurkat cells treated with chemotherapeutic drugs in order to find a new chemotherapeutic target.Methods The expressions of SODD and phospho-NF-?B-p65 proteins in bone marrow cells were detected by immunohistochemistry in 25 children with ALL.The apoptosis incidence was measured by Annexin-V-Fluorescence/PI double-labeling flow cytometry and the expression of SODD and phospho-NF-?B-p65 proteins were determined by Western blotting in Jurkat cells.Results It was found that the expression of SODD and active p65 expression in ALL were significantly higher than those in healthy control group.The expression of SODD and phospho-NF-?B-p65 proteins in the high-risk(HR) group was significantly higher than those in standard-risk(SR) group(Pa

Objective To investigate the anti-apoptotic function of clusterin in LNCaP cell line and the role of clusterin antisense oligodcoxynucleotide(AS-ODN)in TNF-?-induced death of LNCaP cell. Methods The wild type LNCaP(group L),LNCaP transfected with the control vector(group M),LNCaP transfected with full-length clusterin expression vector(group A,ie,study group)were cultured.For the de- tection of cytotoxic effect of TNF-?,MTT and ELISA methods were used to determine the cell proliferation and apoptosis of the 3 clones,and the changes of proliferation and apoptosis in A cell after transfection of clusterin AS-ODN were also assessed.Results MTT method showed that the cell proliferation activity(A value)of groups L,M,and A were 0.84?0.03,0.85?0.04,0.95?0.03,respectively;the difference be- tween groups L and M was not significant(P＞0.05);but compared with group A the cell proliferation activ- ity was significantly lower in groups L and M(P＜0.01 for both).ELISA resuhs showed that the A values of groups L,M,and A were 0.59?0.04,0.62?0.03,0.33?0.04,respectively;the difference between groups L and M was not significant(P＞0.05);but compared with group A,the apoptosis rates were significantly higher in groups L and M(P＜0.01 for both).In group A,A values of cell proliferation activity in subgroups control,AS-ODN,TNF-?,TNF-?+AS-ODN were 1.30?0.03,1.25?0.03,0.99?0.03,0.80?0.03, respectively;the differences between each group were significant(P＜0.05 for all).And the A values of cell apoptosis in the above 4 groups were 0.02?0.00,0.21?0.02,0.63?0.07,1.16?0.04,respectively,the differences between each group were significant(P＜0.01 for all).Conclusions Stable transfection and subsequent expression of clusterin result in resistance to the cytotoxic effect of TNF-?.Transfection with clus- terin AS-ODN enhances cytotoxic effect of TNF-?in A cells.These results suggest that clusterin plays an im- portant role in anti-apoptotic function in LNCaP cell line.

Objective To assess the efficacy of two antibiotic prophylactic regimens in a prospective randomized trial in 1 year for patients undergoing insertion of catheters,and to provide the evidence for uniform consensus existing on the timing,route,and choice of antibiotic.Methods During a period of 12 months,78 patients,who consecutively entered the peritoneal dialysis programme,[45 women and 33 men,mean age (48.2?15.7)years] were included.The prophylactic regimens were a single dose of ceftriaxone (1.0 g) given intravenously 30 minutes before surgery (Group A) and given cefazolin (0.25 g/L) i.p.in the each dialysis bag for 3 days postoperatively (Group B).All operations were performed in one room.The wound was observed every day,and body temperature,Count of white blood corpuscle and type,dialysate were examined every day. Results In Group A and B,none of the patients showed peritonitis or wound infection during the post-operative period (within 10 days).One of 39 patients(2.5%) in the group A,and 2 of 39 patients (5.1%) in the group B had exit site infection (P＞0.05).Conclusions There is no significant difference in the incidence of peritonitis and wound infection between two groups. Prophylactic preoperative single-dose antibiotics intravenously do as well as antibiotics given intraperitoneally for peritoneal dialysis catheter insertion,but is much more convenient.

OBJECTIVETo investigate the differential expression of apoptosis associated gene Bcl-2 and Bax through cell cycle and its possible clinical meaning.

METHODSThe prostate cancer cell line PC-3 was synchronized in M, G1, S and G2 phase using modified thymine deoxyriboside blockage and high pressure N2O technique. The efficiency of synchronization was detected by flow-cytometry. RT-PCR and Western blot methods were used to examine the expression of Bcl-2 and Bax in mRNA and protein level.

RESULTSThe synchronized rate of M, G1, S and G2 phase were 92.1%, 87.0%, 80.2% and 75.9% respectively. Bcl-2 was constitutively expressed through the cell cycle, but both the mRNA and protein expression level of Bcl-2 were very high in the G1 phase, dramatically decreased in M, S and G2 phase. The expression level of Bax had no change through the cell cycle.

CONCLUSIONSCell cycle could influence the expression level of Bcl-2 significantly but not Bax, these might have some clinical relevance.

Cell Cycle ; Cell Line, Tumor ; Gene Expression ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; bcl-2-Associated X Protein ; biosynthesis ; genetics

OBJECTIVETo define changes in clusterin expression following short-term neoadjuvant hormone therapy (NHT) and its biological significance in prostate cancer tissues.

METHODSTwenty-six archival radical prostatectomy (RP) specimens without receiving NHT, 19 needle biopsies and corresponding 19 RP specimens following 3-month NHT, were subjected to immunohistochemical clusterin staining.

RESULTSStaining for clusterin was mainly found in cytoplasm and part of extracellular matrix. Clusterin expression was significantly greater in RP specimens with preoperative NHT (t = 2.91, P < 0.01); Needle biopsies obtained before NHT consistently demonstrated lower staining intensity (1.42 +/- 0.51) than corresponding RP specimens (2.16 +/- 0.60) following 3-month NHT (t = 7.10, P < 0.01).

CONCLUSIONSUpregulation of clusterin in part accounts for malignant progression of prostate cancer through its anti-apoptotic action following androgen withdrawal. These findings support that adjuvant therapy targeting clusterin may enhance androgen ablation therapy in advanced prostate cancer.

Aged ; Clusterin ; genetics ; metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoadjuvant Therapy ; methods ; Oligonucleotides, Antisense ; therapeutic use ; Prostatic Neoplasms ; metabolism ; pathology ; therapy