Objective To observe the influence of lysyl oxidase(LOX)downregulation via RNAi on hypoxic metastasis of human lung cancer cell 95D and stduy its molecular mechanism.Methods LOX siRNA was used to transfect 95D cell line in normoxia (19％ O2 ).After 24-hour incubation,the cells were cultured in hypoxic incubator (0.5％ O2 ) for 24h.Real-time PCR assay was applied to detect LOX mRNA and Snail mRNA expression.Levels of Src,phosphorylation of Src (P-Src Y418 ) and Snail protein were determined by Western blot assay.Transwell chamber was used to evaluate the cellular invasion potential.Results Compared with 95D cells under normoxic conditions,hypoixa treatment increased LOX mRNA expression by 14 times and invasion ability by 2.12 times respectively.Compared with siRNA control group,LOX siRNA transfection decreased LOX mRNA expression,the invasion ability of hypoxic cells,and the protein expression of P-Src Y418 and Snail by 70％ - 75％,about 30％,and about 40％ respectively (P ＜ 0.05).However,it didn't affect the expression level of Src protein or Snail mRNA ( P ＞ 0.05).Conclusions Impaired metastatic potential of hypoxic human lung cancer cell induced by LOX downregulation is associated with reduced expression level of Src activation and Snail protein.The present data provids experimental evidence for LOX as a potential target for prevention and treatment of lung cancer metastasis under hypoxia.

OBJECTIVETo study the dynamic changes of T lymphocyte subsets of AIDS patients during more than 24 months of highly active antiretrovirus therapy (HAART) with successful suppression of HIV replication and different CD4 + T cell restoration.

METHODSTotally 45 AIDS patients who had received HAART for more than 24 months were included. During HAART (including DO, M3, M6, M12, M18, and M24), the number of plasma HIV-1 RNA was measured quantitatively using the bDNA assay, and T lymphocyte subsets including CD3 + CD4 + cells, CD3 + CD8 + cells, naive CD4 + cells (CD4 + CD45RA + CD62L +), CD4 + CD28 + cells , and CD8 + CD38 + cells were detected with flow cytometer.

RESULTSAmong 45 patients, 24 patients (53.3%) whose plasma viral load decreased to less than 500 copies/ml at M6 and maintained to M24 were classified into three groups according to the CD4 + T cell count increments on M24 (compared with DO): group A (< 100/mm3), group B (100-200/mm3), and group C (> 200/mm3). After the initiation of HAART, T lymphocyte response, including CD4 + T cell counts, naive CD4 + cell counts, percentages of CD4 + CD28 + cells in these patients were improved gradually, while CD8 + CD38 + percentage decreased. The improvement of T lymphocyte response in group C was most remarkable even with highest plasma viral load and lowest CD4 T cell count on DO. Compared with group A and B, group C had significantly better improvement not only in the quantities of CD4 + T cell, but also in the CD28 + expression and naive CD4 + T cell populations.

CONCLUSIONST lymphocyte response of AIDS patients can be effectively reconstituted by HAART. Different dynamics of CD4 + CD28 + and naive CD4 + populations may considerably contribute to the quantity and cellular function restoration of CD4 + T lymphocyte.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Anti-HIV Agents ; therapeutic use ; Antiretroviral Therapy, Highly Active ; CD4-Positive T-Lymphocytes ; immunology ; HIV ; drug effects ; physiology ; Humans ; T-Lymphocyte Subsets ; Virus Replication ; drug effects

OBJECTIVETo study the alteration of the expression of CD28 on CD4 + T cells in HIV/AIDS patients and observe the dynamics of CD28 expression under highly active antiretroviral therapy (HAART).

METHODSThe expression of CD28 on CD4 + T cells, CD4 counts, and plasma viral load were measured by flow cytometry and bDNA assays in 278 treatment-naïve HIV/AIDS patients and 56 healthy controls. In addition, the evolution of these parameters was assessed in 59 patients who initiated HAART and were followed for 12 months in regular 3-month visits.

RESULTSThe median level of CD28 on CD4 + T cells decreased dramatically in treatment-naïve HIV-positive individuals than in HIV-negative controls (P <0.001). The expression rate of CD28 molecule was positively correlated with CD4 counts (r = 0.484, P < 0.001), and negatively correlated with plasma viral load (r = -0.300, P <0.001). In patients who had received one month of standard HAART, the level of CD28 on CD4 + T cells increased rapidly from 75.0% to 90.0% (P < 0.001). Moreover, there was a negative correlation between the median CD28 expression and the median viral load (r = - 0.829, P = 0.042).

CONCLUSIONSThe level of CD28 expression on CD4 + T cells is down-regulated in treatment-naïve HIV/AIDS patients. HAART can successfully restore the lymphocyte subsets of CD4 + CD28 + T cells. The up-regulation of CD28 expression after HAART may be closely correlated with the suppression of the viral replication.

Adult ; Antiretroviral Therapy, Highly Active ; CD28 Antigens ; metabolism ; CD4-Positive T-Lymphocytes ; immunology ; Female ; Flow Cytometry ; Follow-Up Studies ; HIV Infections ; blood ; drug therapy ; immunology ; Humans ; Immunologic Memory ; Male ; Middle Aged ; Viral Load

OBJECTIVETo investigate the value of contrast-enhanced ultrasound in the differential diagnosis of solid focal lesions of pancreas (s-FLPs).

METHODSWe retrospectively analyzed the clinical data of 56 s-FLPs examined with contrast agent combined with low mechanical indicators contrast-enhanced ultrasound.

RESULTSThe surrounding pancreas parenchyma enhancement time, lesion enhancement time, and peak enhancement time between different groups of s-FLPs had no significant differences (P > 0.05), while the beginning to peak enhancement time showed significant difference (P < 0.05). When using the enhancement speed as a diagnostic indicator to differentiate pancreatic carcinoma from tumor-like pancreatitis, the sensitivity, specificity, and accuracy were 90.5%, 71.4%, and 85.7% for pancreatic carcinoma and 75.0%, 91.7%, and 89.3% for tumor-like pancreatitis. When using the enhancement pattern as a diagnosis indicator to differentiate pancreatic carcinoma from tumor-like pancreatitis, the sensitivity, specificity, and accuracy were 85.7%, 78.6%, and 83.9% for pancreatic carcinoma and 75.0%, 100%, and 94.6% for tumor-like pancreatitis. When different indicators were combined, enhancement pattern and enhancement speed showed the best diagnostic results; however, the Youden index was not improved.

CONCLUSIONSDifferent s-FLPs show different enhancement findings on contrast-enhanced ultrasound. The enhancement pattern and enhancement speed are the most useful diagnostic indicators.

Diagnosis, Differential ; Humans ; Pancreas ; diagnostic imaging ; Pancreatic Neoplasms ; diagnostic imaging ; Pancreatitis ; diagnostic imaging ; Retrospective Studies ; Ultrasonography

OBJECTIVETo evaluate the clinical performance of an innovative contrast-enhanced ultrasound (CEUS) technique in characterization of focal liver lesions (FLL).

METHODSCEUS was performed in 505 patients with FLLs. A sulfur hexafluoride-filled microbubble ultrasound contrast agent and a continuous real-time imaging technique of contrast pulse sequencing were used. The observation items included initial nodule enhancement time, enhancement level and pattern, and dynamic evolution of enhancement along with the CEUS phases.

RESULTSThere were similar findings for both malignant and benign FLLs in initial enhancement time and enhancement level on CEUS. The dynamic evolution of enhancement along with enhancement phase was the most significant parameter for characterization of FLLs. Most malignant lesions showed enhancement at arterial phase and hypo-enhancing at late phase, whereas the benign lesions presented enhancement at arterial phase and persistent hyper- or iso-enhancing until late phase. Using these characteristics as a diagnostic criteria, the sensitivity, specificity, positive predicative value, negative predicative value and accuracy of CEUS in identifying malignant FLLs were 95.7% (313/327), 86.0% (153/178), 92.6% (313/338), 91.6% (153/167) and 92.3% (466/505), respectively.

CONCLUSIONSCharacterization of FLLs by using the innovative CEUS technique is recommendable due to its safety, facility and high accuracy.

Adult ; Aged ; Aged, 80 and over ; Diagnosis, Differential ; Female ; Humans ; Liver Diseases ; diagnostic imaging ; Male ; Middle Aged ; Sulfur Hexafluoride ; Ultrasonography ; methods

BACKGROUNDHighly active antiretroviral therapy (HAART) produces profound suppression of HIV replication, substantial increase in CD4(+) T cells, and partial reconstitution of the immune system. However, the numbers of subjects were small in previous Chinese studies. This study evaluated the efficacy and side effects of HAART in Chinese advanced AIDS patients.

METHODSOne hundred and three antiretroviral drug naive AIDS patients were enrolled in this study and were divided into two groups by their baseline CD4(+) count: < 100 cells/microl or > or = 100 cells/microl. Clinical, virological and immunological outcomes were monitored at baseline and at 1, 3, 6, 9 and 12 months during the course of treatment with HAART.

RESULTSOne patient died and another was lost from the follow-up. For the remaining 101 HIV/AIDS patients at the 12th month during the HAART, the plasma viral load (VL) was reduced to (3.2 +/- 0.7) lg copies/ml, the CD4(+) count increased to (168 +/- 51) cells/microl [among which the naive phenotype (CD45RA(+)CD62L(+)) increased to (49 +/- 27) cells/microl and the memory phenotype (CD45RA(-)) increased to (119 +/- 55) cells/microl], and the percentage of CD4(+)CD28(+) cells increased. At the same time, there was a significant reduction of CD8(+) T cell activation. In the 69 patients with the baseline CD4(+) count < 100 cells/microl, 37 had a VL < 50 copies/ml; while in the 34 patients with the baseline CD4(+) count > or = 100 cells/microl, 25 had a VL < 50 copies/ml, the difference between the two groups was statistically significant. The CD4(+) T cell count showed a two-phase increase during HAART and a significant positive correlation was shown between the change of CD4(+) count and plasma VL. Over 12 months of HAART, 10 patients had gastrointestinal side effects, 13 peripheral neuritis, 7 hepatic lesions, 8 hematological side effects, 8 skin rashes, 10 lipodystrophy and 1 renal calculus.

CONCLUSIONSImmune reconstitution as well as the significantly improved clinical outcomes is observed in Chinese advanced AIDS patients after HAART. Side effects are common during HAART and require clinical attention.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; virology ; Adult ; Antiretroviral Therapy, Highly Active ; CD28 Antigens ; analysis ; CD4 Lymphocyte Count ; Female ; Humans ; Male ; Middle Aged ; RNA, Viral ; blood ; Viral Load

BACKGROUNDThe correlation between HIV-1 Nef-specific CD8 T-cell responses and markers of HIV-1 disease progression still remains unclear. This study analysed and compared the role of HIV-1 Nef-specific CD8 T-cell responses in patients with different disease status.

METHODSTwo groups of patients with HIV-1 subtype B infection were selected according to CD4 count and clinical manifestations: long-term nonprogressors (LTNPs, n = 20) and advanced progressors (APs, CD4 count < 500 cells/microl, n = 34). Nef-specific CD8 T-cell responses were studied by interferon-gamma ELISpot assay against 3 pools of HIV-Nef peptides.

RESULTSNef-specific CD8 T-cell responses did not correlate with viral load or CD4 count in all patients and no significant differences were found in the magnitude of Nef-specific CD8 T-cell responses between groups LTNPs and APs (670 SFC/10(6) peripheral blood mononuclear cells vs 1107 SFC/10(6) peripheral blood mononuclear cells, P = 0.255). Further comparisons showed that there were also no significant correlations observed in group LTNPs, but Nef-specific CD8 T cells correlated negatively with viral load (r = -0.397, P = 0.020) and positively with CD4 count (r = 0.364, P = 0.034) in group APs.

CONCLUSIONThese data suggest that different correlation patterns between Nef-specific CD8 T-cell responses and disease progression exist in LTNPs and APs. Although a negative association was observed with concurrent plasma HIV RNA in APs, Nef-specific CD8 T-cell responses might fail to play a protective role in different stages of HIV-1 infection.

Acquired Immunodeficiency Syndrome ; immunology ; Adult ; CD4 Lymphocyte Count ; CD8-Positive T-Lymphocytes ; immunology ; Disease Progression ; Female ; Gene Products, nef ; immunology ; HIV-1 ; classification ; Humans ; Male ; Middle Aged ; RNA, Viral ; blood ; nef Gene Products, Human Immunodeficiency Virus

Suppression of cellular O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) can repress prolifera-tion and migration of various cancer cells,which opens a new avenue for cancer therapy.Based on the regulation of insulin gene transcription,we designed a cell-based fluorescent reporter capable of sensing cellular O-GlcNAcylation in HEK293T cells.The fluorescent reporter mainly consists of a reporter (green fluorescent protein (GFP)),an internal reference (red fluorescent protein),and an operator (neuronal differentiation 1),which serves as a "sweet switch" to control GFP expression in response to cellular O-GlcNAcylation changes.The fluorescent reporter can efficiently sense reduced levels of cellular O-GlcNAcylation in several cell lines.Using the fluorescent reporter,we screened 120 natural products and obtained one compound,sesamin,which could markedly inhibit protein O-GlcNAcylation in HeLa and human colorectal carcinoma-116 cells and repress their migration in vitro.Altogether,the present study demonstrated the development of a novel strategy for anti-tumor drug screening,as well as for con-ducting gene transcription studies.

OBJECTIVETo investigate the clinical characteristics of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients in China.

METHODSTotally 143 HIV/AIDS patients who were first diagnosed in Peking Union Medical College Hospital form January 1988 to April 2006 were enrolled in this study. Clinical characteristics were retrospectively analyzed.

RESULTSAmong 143 HIV/ AIDS patients, 57 patients had no clinical symptoms and were confirmed by routine examinations; 86 patients had clinical symptoms, including fever (n = 50), weight loss (n = 18), and discomforts involving respiratory system (n = 34), gastrointestinal system (n = 16), and derma and mucosa (n = 17). Opportunistic infections (OIs) such as pneumocystis jiroveci pneumonia (PCP) (n = 27), oropharyngeal candidiasis (n = 16), tuberculosis (n = 15) , and cytomegalovirus (CMV) infection (n = 9) were also observed in patients whose CD4 + T cell counts were less than 200/mm3. Most CMV infection and cryptococcal meningitis occurred in patients whose CD4 + T cell counts were less than 100/mm3. CD4 + T cell count was negatively correlated with plasma viral load (r = -0.420, P = 0.001).

CONCLUSIONSFever, dyspnea, and weight loss are the most common symptoms in the patients of this study. The respiratory system, gastrointestinal system, derma and mucosa are the most commonly affected areas by OIs, and PCP is the most common OI. The occurrence of OIs corelates with CD4 + T cell count.

AIDS-Related Opportunistic Infections ; immunology ; Acquired Immunodeficiency Syndrome ; complications ; Adolescent ; Adult ; Aged ; CD4 Lymphocyte Count ; China ; Dyspnea ; etiology ; Emaciation ; etiology ; Female ; Fever ; etiology ; HIV Infections ; complications ; Humans ; Male ; Middle Aged ; Pneumonia, Pneumocystis ; immunology ; Retrospective Studies

BACKGROUNDThe conventional approaches to diabetes screening are potentially limited by poor compliance and laboratory demand. This study aimed to evaluate the performance of fasting plasma glucose (FPG) and postprandial urine glucose (PUG) in screening for diabetes in Chinese high-risk population.

METHODSNine hundred and nine subjects with high-risk factors of diabetes underwent oral glucose tolerance test after an overnight fast. FPG, hemoglobin A1c, 2-h plasma glucose (2 h-PG), and 2 h-PUG were evaluated. Diabetes and prediabetes were defined by the American Diabetes Association criteria. The area under the receiver operating characteristic (ROC) curve was used to evaluate the diagnostic accuracy of 2 h-PUG, and the optimal cut-off determined to provide the largest Youden index. Spearman correlation was used for relationship analysis.

RESULTSAmong 909 subjects, 33.4% (304/909) of subjects had prediabetes, and 17.2% (156/909) had diabetes. The 2 h-PUG was positively related to FPG and 2 h-PG (r = 0.428 and 0.551, respectively, both P < 0.001). For estimation of 2 h-PG ≥ 7.8 mmol/L and 2 h-PG ≥ 11.1 mmol/L using 2 h-PUG, the area under the ROC curve were 0.772 (95% confidence interval [CI ]: 0.738-0.806) and 0.885 (95% CI: 0.850-0.921), respectively. The corresponding optimal cut-offs for 2 h-PUG were 5.6 mmol/L and 7.5 mmol/L, respectively. Compared with FPG alone, FPG combined with 2 h-PUG had a higher sensitivity for detecting glucose abnormalities (84.1% vs. 73.7%, P < 0.001) and diabetes (82.7% vs. 48.1%, P < 0.001).

CONCLUSIONFPG combined with 2 h-PUG substantially improves the sensitivity in detecting prediabetes and diabetes relative to FPG alone, and may represent an efficient layperson-oriented diabetes screening method.

Aged ; Asian Continental Ancestry Group ; Blood Glucose ; metabolism ; Diabetes Mellitus ; blood ; diagnosis ; urine ; Fasting ; blood ; Female ; Glucose Tolerance Test ; Humans ; Male ; Mass Screening ; methods ; Middle Aged ; Postprandial Period ; physiology