The polyclonal antibodies against VLDL receptor were prepared and identified. Rabbits were immunized with polypeptide fragment of VLDL receptor as antigen. The collected blood serum of the immunized rabbits was analyzed and identified by using ELISA and Western Blot. The results showed that the rabbit against mouse and human VLDL receptor antibodies were obtained with high titer and could recognize the natural VLDL receptors through Western blot. The prepared polyclonal antibodies against VLDL receptor provide a new tool to study the protein of VLDL receptor.
Antibodies/chemistry ; Antibodies/*immunology ; Enzyme-Linked Immunosorbent Assay ; Peptides/*immunology ; Receptors, LDL/*immunology

OBJECTIVESTo analyze the relationship between oxidized low density lipoprotein (oxLDL), angiogenesis and stabilization of atherosclerotic plaques in human coronary arteries; and to investigate the role of oxLDL in creating vulnerable sites in atherosclerotic plaques.

METHODSSamples of coronary arteries were obtained at autopsies of 42 patients with acute coronary syndrome. Eighty randomly selected blocks were studied by immunohistochemistry using antibodies against oxLDL and endothelial cells (factor VIII). Computer-aided planimeter was used for quantitative analysis.

RESULTSIn unstable plaques, percentage of immunoreactive areas for oxLDL was significantly higher than that in stable plaques. Most of the oxLDL were located in shoulder region of these plaques, as compared to the fibrous cap and basal regions. The details of distribution of oxLDL were as follows: shoulder region (20.43 +/- 3.12 for unstable plaques and 17.65 +/- 4.22 for stable plaques), fibrous cap (4.77 +/- 2.03 for unstable plaque and 2.80 +/- 0.22 for stable plaques) and basal region (5.65 +/- 1.65 for unstable plaques and 3.22 +/- 1.02 for unstable plaques). OxLDL was also a main component in the lipid core. In the shoulder region, there was a significant positive correlation between neovascularization and oxLDL (r = 0.8247, P = 0.000).

CONCLUSIONSThe amount of oxLDL is significantly higher in unstable atherosclerotic plaques, especially over the shoulder region. OxLDL in coronary atherosclerotic plaques is thus an important factor in determining stabilization of the plaques. OxLDL may induce influx of inflammatory cells which subsequently leads to decreased plaque stabilization.

Angina, Unstable ; metabolism ; pathology ; Coronary Artery Disease ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Lipoproteins, LDL ; metabolism ; Myocardial Infarction ; metabolism ; pathology ; Neovascularization, Pathologic ; metabolism ; pathology

OBJECTIVETo compare the angiogenesis in unstable and stable plaques and to investigate the potential role of neovessels in creating vulnerable sites for atherosclerotic plaques.

METHODSSpecimens of coronary arteries were obtained from 52 autopsy cases with acute coronary syndromes. Plaque morphology was studied by use of stained slides. 922 tissue blocks of late-stage lesions were classified into two groups: (1) unstable plaque (n = 153), the plaque was characterized by a large extracellular lipid core (more than 40% of the plaque area); (2) stable plaque (n = 769), lipid core less than 40% of the plaque area. Forty blocks were selected randomly from each group and serial sections were stained immunohistochemically with a polyclonal antibody against F VIII RAg. Computer-aided planimeter was used for quantitative analysis.

RESULTSIn unstable plaques, the occurrence of neovessels was more frequent and the neovessel density (number/mm(2)) was significantly increased as compared to that of stable plaques (frequency: 80.4% vs 66.6%, P < 0.01; shoulder: 22.16 +/- 19.96 vs 10.04 +/- 11.52, base: 21.68 +/- 20.44 vs 9.68 +/- 11.52, fibrous cap: 3.80 +/- 5.32 vs 1.48 +/- 2.28, P < 0.05). Most neovessels were located in the shoulder region and at the base of plaques.

CONCLUSIONSThese findings suggest that neovessels in coronary atherosclerotic plaques are closely associated with the decreased stabilization of the plaques.

Aged ; Aged, 80 and over ; Coronary Artery Disease ; pathology ; Coronary Vessels ; pathology ; Female ; Humans ; Male ; Neovascularization, Pathologic ; pathology

The polyclonal antibodies against VLDL receptor were prepared and identified. Rabbits were immunized with polypeptide fragment of VLDL receptor as antigen. The collected blood serum of the immunized rabbits was analyzed and identified by using ELISA and Western Blot. The results showed that the rabbit against mouse and human VLDL receptor antibodies were obtained with high titer and could recognize the natural VLDL receptors through Western blot. The prepared polyclonal antibodies against VLDL receptor provide a new tool to study the protein of VLDL receptor.
Animals ; Antibodies ; chemistry ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Peptides ; immunology ; Rabbits ; Receptors, LDL ; immunology

OBJECTIVETo investigate the difference of the antigenicity of xenogenic acellular dermal matrix (ADM) implants prepared by different methods.

METHODSThe split-thickness skin sheet from swine was processed by trypsin and Triton X-100 to make xeno-ADM. Twenty-five Japanese white rabbits were divided into 5 groups, i.e. xeno-ADM(1) (conjugated with glutaraldehyde), xeno-ADM(2) (conjugated with network) and xeno-ADM(3) (no conjugation, as control), in which the ADMs were and xeno-ADM(4) (conjugated) and allo-ADM (no conjugated as control), in which the ADMs were embedded into the subcutaneous place of rabbit ear and back after that the rabbits were pre-sensitized by xeno-ADM soluble protein antigen injections. The titers of anti ADMs antibody in rabbit serum were monitored during 2 - 32 post-operative weeks and the histological changes of the embedded ADMs were observed grossly and microscopically.

RESULTSThe serum titers of anti-xeno-ADM in xeno-ADM(4) group was the highest. Whereas regardless of the sensitizing effects, the titers in all groups ranged as follows: xeno-ADM(3) > xeno-ADM(2) > xeno-ADM(1) (P < 0.05 - 0.01). About 40% serum samples in allo-ADM group exhibited positive anti-allo-ADM protein antibodies. Histologically, Evident and lasting inflammatory reaction could be found in the xeno-ADM grafting sites, which was much stronger than that in allo-ADM group. The degradation and absorption gradient of ADM was ranked as follow: xeno-ADM(3) > xeno-ADM(2) > xeno-ADM(4) > xeno-ADM(1) > Allo-ADM. Foreign body megalocytic reaction might evoke in the surrounding of conjugated ADM.

CONCLUSIONThe immunogenicity in xeno-ADM was stronger than that in allo-ADM, which could induce the host to develop immune reaction restricted by IgG. Large sheets of degenerated ADM implants could lower down the antigen-antibody reaction and ameliorate the structural destroying and degeneration absorption of ADM induced by inflammatory immune reaction.

Animals ; Antigen-Antibody Reactions ; Dermis ; immunology ; transplantation ; Male ; Rabbits ; Skin Transplantation ; Swine ; Transplantation, Heterologous

Objective To probe into the way for relieving pains irrelevant to incision but associated with the event of pneumoperitoneum by means of physiotherapy, wherein the said pains occur after a gynecologic celioseope operation.Methods Random option of 207 patients after the gynecologic operation was conducted and the patients were divided into 3 groups at random that were on the man-care,instrument and ordinary basis.Each group consists of 69 cases.In man-care group,artificial massage at the points (hereafter called mann-care group)Was adopted.The instrument group adopted microwave devices to heal (hereafter called instrument group).And the ordinary group adopted regular care of post operation (hereafter called ordinary group).We observed and compared three methods,to see whether they may relieve the pains irrelevant to incision.Results The point massage and microwave therapy can relieve the complicated pains irrelevant to incision after a gynecologic operation,and the effect thereof is obviously better than that of the ordinary care.Conclusions As to the complications such as pains irrelevant to incision after gynecologic operation,the point massage,microwave and relevant physiotherapy can function well and are comparatively ideal.

Objective To investigate the effects of different culture conditions(RPMI-1640,DMEM and DMEM/F12 medium)on the passage of MPM cells isolated from the tissues of Malignant pleural mesothelioma(MPM),and to study the effects of CDKN2B on the proliferation,invasion and apoptosis of MPM cells.Methods MPM cells were isolated from MPM tissues and cultured in RPMI-1640,DMEM and DMEM/F12 medium,respectively.Cell proliferation was examined by CCK-8,and the nuclei and chromosomes were observed by Wright-Giemsa staining.Fluorescence intensities of Calretinin,CD141,CK5,EMA and WT-1 were conducted by immunofluorescence assay.The mRNA and protein expression of CDKN2B were detected by RT-qPCR and Western blot,respectively.Transwell was used to detect cell invasion and flow cytometry was used to detect cell apoptosis.Results The established MPM cells showed good viability when passaged to the 10th generation in RPMI-1640,DMEM and DMEM/F12 cultures,and the MPM markers Calretinin,CD141,CK5,EMA and WT-1 were all expressed in the cells.The viability of MPM cells in RPMI-1640 culture medium was relatively stable.CDKN2B was downregulated in MPM cells(P<0.05),and overexpression of CDKN2B significantly suppressed the proliferation(P<0.05),invasion(P<0.05)and epithelial interstitial transformation of MPM cells(P<0.01),and promoted the apoptosis(P<0.01).Conclusion The established MPM cells were stably passaged in RPMI-1640 culture medium,and CDKN2B may be a potential target for the diagnosis and treatment of MPM.

OBJECTIVETo explore an ideal skin substitute with its appearance and texture similar to normal skin, to repair wounds with full-thickness skin defect.

METHODSComposite skin (CS) in question was composed of allo/xenogeneic acellular dermal matrix (ADM) and razor thin autoskin. One step skin grafting was employed in the experimental study and clinical trial. Razor thin autoskin alone was used as the control in the study. Changes in the antigenicity of ADM and the reformation of basement membrane (BM) structure at epidermis-dermis junction (EDJ) of ADMs were studied at designated time points after the grafting with biochemical and immunohistochemical methods. Fifty-three patients with full thickness skin defects due to various causes, including scar excision were grafted with CS, and survival rate and long-term result were observed.

RESULTSThe grafted CS survived satisfactory. The reformation of the basement membrane structure was clearly observed at the 28th post-graft week. The basement membrane cells grew with polarization in an undulating arrangement. There was reformation of dermal papillae and ridges. The antigenicity of allo-ADM was obviously lower than that of xeno-ADM. Sixty-five out of 70 pieces of CS grafting (92.9%) survived totally, two of them survived partially, and three failed due to infection. The longest follow-up period was 8 and a half years. The grafted CS appeared similar to the normal skin in regard to the texture and color, especially allo-ADM, and no evident rejection reaction was seen.

CONCLUSIONADM possessed very low antigenicity, thus serving a lasting framework after grafting. In addition, it could serve as a "dermal template" for the induction of tissue regeneration.

Animals ; Burns ; surgery ; Dermis ; transplantation ; Follow-Up Studies ; Graft Survival ; Humans ; Male ; Rabbits ; Skin Transplantation ; methods ; Swine ; Transplantation, Autologous ; Transplantation, Heterologous ; Transplantation, Homologous ; Treatment Outcome ; Wound Healing

OBJECTIVETo investigate the role of postoperative chemoradiotherapy (CRT) as a multimodality treatment option for locally advanced thoracic esophageal squamous cell carcinoma (ESCC) by a prospective comparison between surgery alone and postoperative CRT.

METHODSUsing preoperative computed tomography (CT)-based staging criteria, 158 patients with ESCC (stage II-III) were enrolled in this prospective study. With informed consent, the patients were randomized into two groups: postoperative CRT (78 cases) and surgery alone (S, 80 cases). After a few minor adjustments to the enrolled patients, the actual patients of postoperative CRT group and S group were 74 cases and 77 cases, respectively. Comparison of the complications, local recurrence rate, distant metastasis rate, survival rate and progression-free survival in the two groups was carried out.

RESULTSWith a median follow-up of 37.5 months, the 1-, 3-, 5-, 10-year overall survival (OS) rates were 91.0%, 62.8%, 42.3%, 24.4% and 87.5%, 51.3%, 33.8%, 12.5% for the postoperative CRT and S arm, respectively. A significant difference in OS was detected between the two arms (P = 0.0276). There was a significant difference of progression-free survival (PFS) between the two arms (P = 0.0136). The local recurrence rates in the postoperative CRT group and S group were 14.9% and 36.4%, respectively (P < 0.05). No significant difference was detected between the complications of the two groups (P > 0.05). Toxicities of chemoradiotherapy in the postoperative CRT arm were moderate, which can be relieved rapidly by adequate therapy.

CONCLUSIONRational application of postoperative chemoradiotherapy can provide a benefit in progression-free survival and overall survival in patients with locally advanced esophageal squamous cell carcinoma.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Squamous Cell ; pathology ; therapy ; Chemotherapy, Adjuvant ; Cisplatin ; administration & dosage ; Disease-Free Survival ; Esophageal Neoplasms ; pathology ; therapy ; Esophagectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Lymph Node Excision ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Paclitaxel ; administration & dosage ; Prospective Studies ; Radiotherapy, Adjuvant ; Radiotherapy, High-Energy ; Survival Rate

BACKGROUND: The X-ray 9-point grading system is established based on cervical disc degeneration and its suitability for evaluation of lumbar disc degeneration has not been validated. OBJECTIVE: To investigate the feasibility of the evaluation of lumbar intervertebral disc degeneration using the 9-point grading system with X-ray, and to explore its clinical application value. METHODS: The correlation between 9-point grading system and Pfirrmann system was statistically analyzed. Magnetic resonance imaging (MRI) and X-ray images of 172 cases with lumbar disc degeneration or simple lumbago, who were treated in the Shanghai Changzheng Hospital from June 2013 to June 2016, were retrospectively analyzed. The MRI scores of lumbar disc degeneration were evaluated according to Pfirrmann standard, and the X-ray scores were evaluated according to 9-point grading system. The two groups of scores were respectively analyzed by the correlation and regression tests via Spearman rank sum test with SPSS software. RESULTS AND CONCLUSION: (1) MRI scores (the independent variable X): 2 cases scored 1; 8 cases scored 2; 55 cases scored 3; 84 cases scored 4; 23 cases scored 5. X-ray scores (the dependent variable Y): 2 cases scored 0; 6 cases scored 1; 21 cases scored 2; 43 cases scored 3; 40 cases scored 4; 26 cases scored 5; 16 cases scored 6; 8 cases scored 7; 7 cases scored 8; 3 cases scored 9. (2) The Spearman correlation test showed that significant positive correlation was found between Pfirrmann system and 9-point grading system (P <0.01). Significance test of correlation coefficient proved that there was significant linear correlation between Pfirrmann system and 9-point grading system. Further regression analysis showed the linear regression between Pfirrmann system and 9-point grading system. (3) These findings suggested that Pfirrmann system and X-ray 9-point grading system had a good consistency in evaluating lumbar disc degeneration. As a scientific, fast and effective evaluation method, the X-ray 9-point grading system has a certain reference value in the diagnoses of lumbar degenerative diseases.