p53 gene mutations and their expression in protein levels of colorectal adenomas, carcinomas and regional lymph nodes metastasis were detected by PCR-SSCP and AB-PAP immunohistochemical analysis, respectively. The results showed that p53 gene mutations were detected in 25% colorectal adenomas, 80% carcinomas and 100% regional lymph nodes metastasis. Meanwhile, p53 proteins were detected at rates of 75%, 60% and 57% in these three groups, respectively. It is suggested that p53 gene mutations occur frequently at late stage of progression to colorectal adenoma-carcinoma sequence and may relate to its metastasis, while the accumulation of p53 protein may be a specific genetic alteration initiated from adenomatous stage. Thus, the biological significances of the two genetic alterations are different. Investigations of these concomitantly would be helpful in understanding the malignant potential of adenoma and evaluation of its staging.

Objective To observe the cellular phenotype conversion of human mesenchymal stem cells (MSCs) cocultured indirectly with heat-shocked human sweat gland cells (SGCs) in vitro and explore the relative mechanism. Methods MSCs and SGCs were isolated and amplified in vitro. First,primary confluent cultures of SGCs were heat-shocked at 47℃. Then, the supernatants were collected immediately and 24 hours before applied to the third generation of MSCs. After seven days, the MSCs expressing CK7, CK18 and CEA were examined by two-step immunocytochemistry and flow cytometry and compared with the control group. Results MSCs treated with the supernatants of SGCs proliferated slowly, with no obvious morphological changes during seven days. Two-step immunocytochemistry demonstrated positive staining of CK7 and CEA in some cells. Additionally, the positive rate of CK7 and CEA was 5.76% and 2.01% by flow cytometry, much higher than that of the control sample, which was only 1.12% and 0.51% respectively (P ＜ 0.01 ). Conclusions There are some signal moleculars in the supernatants of heat-shocked SGCs, which benefits the transdifferentiation of MSCs.

Objective To evaluate the protective effects of tea polyphenols against the destruction of melanocytes by CD8+ T cells from vitiligo patients.Methods Skin tissue was resected from the margin of vitiligo lesions followed by the isolation and culture of CD8+ T lymphocytes,and from the normal skin of vitiligo patients followed by the isolation and culture of melanocytes.Flow cytometry was carried out to evaluate the purity of CD8+ T cells.The melanocytes were cocultured with the CD8 + T cells at different ratios followed by the evaluation of killing effect of CD8+ T cells.Various concentrations (200 and 400 μg/ml) of tea polyphenols were added into the co-culture system of CD8+ T cells and melanocytes at a ratio of 5 ∶ 1 followed by an additional culture of 48 hours.Then,flow cytometry was performed to detect the apoptosis in melanocytes in the coculture system.Results CD8+ T lymphocytes were successfully obtained from the marginal area of vitiligo lesions with a purity of more than 90％,which highly expressed the antigens CD137 and CD69.The coculture with CD8+ T cells markedly accelerated the apoptosis in melanocytes,while the accelerative effect was inhibited by tea polyphenols of 200 and 400 μg/ml.Conclusions The CD8+ T cells infiltrating the edge of vitiligo lesions display a potential destructive effect on autologous melanocytes from vitiligo patients,and tea polyphenols have a protective effect against the destruction of melanocytes by CD8+ T cells.

Objective To study the influence of UPPP on resonance characteristics of four resonance cavity in patients with OSAHS .Methods Using multi-channel voice analysis system to test the cavity resonance character-istics and energy distribution of 36 OSAHS patients (including pre - and post -operative) and 36 normal males when they speak /a:/.The frequency spectrum was 4 000Hz(FR1 ,FR2 ,FR3 ,FR4 ) .Results FR2 energy values of the head cavity and mouth cavity of OSAHS patients heighten significantly after UPPP (P<0 .05);FR1 energy val-ues of head ,mouth and thoracic cavity were higher than that of in normal males'after UPPP(P<0 .05) .FR2 ,FR3 , FR4 energy values of head cavity ,FR2 ,FR3 energy values of mouth cavity ,FR2 energy values of thoracic cavity were lower than those of in normal males'after UPPP(P<0 .05) .Conclusion UPPP surgery removes the obstruction of the upper airway of OSAHS patients and changes the morphology of the pharynx ,thas changes the resonance charac-teristics of the head cavity and oral .

Objective:To explore the diagnostic value of 64‐slice spiral CT for in‐stent restenosis (ISR) after percuta‐neous coronary intervention (PCI) in patients with coronary heart disease (CHD) .Methods :A total of 120 CHD patients after PCI received 64‐slice spiral CT angiography and routine coronary angiography (CAG ) respectively . Then coronary ISR was assessed .Results:With CAG as the gold standard ,sensitivity ,specificity ,positive predictive value and negative predictive value were 87.5% ,95.3% ,80.0% and 97.3% respectively for 64‐slice spiral CT cor‐onary angiography in diagnosis of ISR .Conclusion :The 64‐slice spiral CT coronary angiography possesses high sensitivity and accuracy diagnosing coronary in‐stent restenosis ,which can be used as one of noninvasive measures for postoperative follow‐up after percutaneous coronary intervention .

Objective To evaluate the effect of calcipotriol on the proliferation of and cytokine secretion by melanocytes and perilesional CD8+ cytotoxic T lymphocytes (CTLs) from patients with vitiligo.Methods Melanocytes isolated from abdominal skin and CD8+ CTLs from perilesional skin of patients with vitiligo were subjected to successive culture in vitro.After several passages,the melanocytes and CD8+ CTLs were cultured alone or in combination with or without the presence of various concentrations of calcipotriol for 24 to 48 hours.MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium,inner salt) method was used to evaluate the proliferative activity of cells,enzyme-linked immunosorbent assay to determine the levels of interleukin (IL)-6,tumor necrosis factor (TNF)-α and interferon (IFN)-γ in the culture supematant of cells,flow cytometry to detect cell apoptosis.Some co-cultured melanocytes and CTLs were treated with calcipotriol of 10-8 mol/L and anti-IL-6 antibody of various concentrations (0,1,2,2.5,5,10 mg/L) for two days followed by enumeration of cells.The concentrations of 108 and 10-9 mol/L (calcipotriol) were chosen for relevant tests.Results There was a marked apoptosis in MCs after coculture with CD8+ CTLs.The 24-hour treatment with calcipotriol of 104 and 10-9 mol/L had no obvious effect on the proliferation of melanocytes cultured alone (both P ＞ 0.05),but accelerated the proliferation of melanocytes cocultured with CTLs (both P ＜0.05) as well as that of CD8+ CTLs cultured alone or in combination with melanocytes (all P ＜0.05).A statistical decrease was observed in IL-6,TNF-α and IFN-γlevels in the supernatant of cocultured melanocytes and CTLs compared with those in the supernatant of melanocytes and CTLs cultured alone,and calcipotriol of 10-9 mol/L intensified the decrease in supernatant IL-6 level (t =2.89,P ＜0.05),but no statistical changes were noted for the level of TNF-α or IFN-γin the supernatant of the coculture system after treatment with calcipotriol of 104 or 104 mol/L compared with that before treatment (both P ＞ 0.05).In the coculture system pretreated with calcipotriol of 10-8 mol/L,the number of CD8+ CTLs significantly decreased (t =3.15,P ＜0.05),whereas that of melanocytes significantly increased (t =3.53,P ＜0.05) after the treatment with anti-IL-6 antibody of 5 mg/L.Conclusions Perilesional CD8+ CTLs have a specific cytotoxic effect on melanocytes,and calcipotriol may inhibit the cytotoxic effect of CD8+ CTLs by suppressing the secretion of IL-6,which may partly explain the therapeutic mechanism of calcipotriol for vitiligo.

Kaposi's sarcoma-associated herpesvirus (KSHV) is causally related to Kaposi's sarcoma (KS),primary effusion lymphoma (PEL) and a proportion of cases of multicentric Castleman's disease (MCD). The ORF73 protein was cloned into pQE80L-orf73 and expressed in E.coli and purified. The expressed recombinant ORF73 was identified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). A protein of about 27 kDa was expressed as expected. Western Blotting showed that the purified recombinant ORF73 reacted with KSHV positive serum. The immunogenicity of the recombinant ORF73 was further analysed by ELISA and the optimal conditions were determined. The ORF73 ELISA was used to compare the KSHV seroprevalence between Hubei and Xinjiang Han people. The Hart people in Xinjiang have significantly higher KSHV seroprevalence than their counterparts in Hubei (6.7% vs 2.9%, P = 0.005).

The ORFK8.1 of Kaposi's sarcoma associated-herpesvirus (KSHV) was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf K8.1 was induced by isopropyl-b-D-thiogalactopyranoside (IPTG). The fusion protein was purified by chromatyography. The expressed protein and its purified product were identified by sodium dodecyl sulfate-polyacrylamide gel eletrophoresis (SDS-PAGE). SDS-PAGE showed that a protein of 26 kDa was visualized as expected. A western blot assay was established to analyze the immunogenicity of purified recombinant 0RFK8.1 protein. The optimal condition of the recombinant ORFK8.1 ELISA assay was confirmed: the concentration of antigen was 5 ug/mL, the dilution of serum was 1:200. We used the ELISA method to investigate the recombinant ORF K8.1 protein's specificity, the data showed that the specificity of ORF K8.1 to detect KSHV was 100%. At the same time, 560 sera samples from Hubei province were detected by using ORFK8.1 ELISA to investigate KSHV seroprevalence in this region. The KSHV seroprevalence in Hubei province is shown to be 6.80%.

Objective To investigate the awareness of osteoporosis and its influential factors in middle-aged and older people. Methods By using self-designed questionnaire, 354 middle-aged and old people were investigated as to their awarness of osteoporosis in Chajiao street community in Guangzhou. Among them, 58 people were measured the bone density in the calcaneal bone by Ultrasound. Results The average score of the questionnaire was 9.46 ± 5.13, and the accuracy was 39.4%. In the diagnosis and treatment, the accuracy was just 17.8%. The diet (the accuracy in 54%) and sport (the accuracy in 50.3%) were good. The statistical analyses showed the linear relationship between the T value of bone density and the age (P = 0.021), between the T value and the score (P = 0.047). Wilcoxon rank sum test found that the T value between female and male was significantly different (Z = -3.749, P < 0.000). With multiple linear regression analysis, the T value had a linear correlation with the score (F = 4.224, P = 0.045) positively. Conclusions Currently, the awareness of osteoporosis in the middle-aged and old people was inadequate. To better prevent osteoporosis, the propogation of knowledge and guidance provided by clinicians in osteoporosis should be improved necessarily and emergently.

BACKGROUND:Cytoskeleton plays an important role in the transduction of mechanical signal, and intermittent tensile stress can promote osteogenic differentiation. However, there is no relevant study about the change of cytoskeleton in osteoporosis rat bone marrow mesenchymal stem cells under intermittent tensile stress. OBJECTIVE:To investigate the effects of intermittent tensile stress on the cytoskeleton of osteoporosis rat bone marrow mesenchymal stem cells during osteogenic differentiation. METHODS:Bone marrow mesenchymal stem cells were obtained from osteoporosis rats and cultured in vitro. The 5%, 10%and 15%tensile stress were strained on the bone marrow mesenchymal stem cells through FX-4000T Flexcell. No stress was in the control group. Osteogenic differentiation of bone marrow mesenchymal stem cells was observed through alkaline phosphatase staining, while the change of cytoskeleton was observed by confocal laser scanning microscopy with figures col ected for analysis by Image-ProPlus 6.0 software. The area of cells, ratio of length to width and integrated fluorescence intensity of cytoskeleton protein F-actin were measured. RESULTS AND CONCLUSION:Under tensile stress, bone marrow mesenchymal stem cells from osteoporosis rats arranged in the direction vertical to mechanical stimulation. cells under different tensile stress differentiated towards osteoblasts. The result of alkaline phosphatase staining showed the most significant difference in 10%group, and quite an amount of cells lining lost succession in the 15%group. Under stress, the F-actin filaments were rearranged in paral el accordingly, which showed a reconstruction of cytoskeleton. Imaging analysis indicated that the area of bone marrow mesenchymal stem cells was decreased in 10%and 15%groups (P<0.05) with the increased ratio of length to width (P<0.05), and expression of F-actin increased in5%, 10%, 15%groups (P<0.05) after tensile stress. Under mechanical stimulation, the cytoskeleton of bone marrow mesenchymal stem cells from osteoporosis rats is shown to have corresponding alterations during osteogenic differentiation.