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The serine/threonine kinase mammalian/mechanistic target of rapamycin (mTOR) integrates various environmental cues such as the presence of antigen, inflammation, and nutrients to regulate T cell growth, metabolism, and function. The tuberous sclerosis 1 (TSC1)/TSC2 complex negatively regulates the activity of an mTOR-containing multiprotein complex called mTOR complex 1. Recent studies have revealed an essential cell-intrinsic role for TSC1 in T cell survival, quiescence, and mitochondrial homeostasis. Given the emerging role of mTOR activity in the regulation of the quantity and quality of CD8 T cell responses, in this study, we examine the role of its suppressor, TSC1, in the regulation of antigen-specific primary and memory CD8 T cell responses to bacterial infection. Using an established model system of transgenic CD8 cell adoptive transfer and challenge with Listeria monocytogenes expressing a cognate antigen, we found that TSC1 deficiency impairs antigen-specific CD8 T cell responses, resulting in weak expansion, exaggerated contraction, and poor memory generation. Poor expansion of TSC1-deficient cells was associated with defects in survival and proliferation in vivo , while enhanced contraction was correlated with an increased ratio of short-lived effectors to memory precursors in the effector cell population. This perturbation of effector-memory differentiation was concomitant with decreased expression of eomesodermin among activated TSC1 knockout cells. Upon competitive adoptive transfer with wild-type counterparts and antigen rechallenge, TSC1-deficient memory cells showed moderate defects in expansion but not cytokine production. Taken together, these findings provide direct evidence of a CD8 T cell-intrinsic role for TSC1 in the regulation of antigen-specific primary and memory responses.

Aims ATP-binding cassette transporter A1 (ABCA1) mediates the efflux of cholesterol and phospholipids to lipid-poor apolipoproteins, which then form nascent HDL, a key step in the mechanism of reverse cholesterol transport (RCT). While a series of microRNAs (miRNAs) have been identified as potent post-transcriptional regulators of lipid metabolism, their effects on ABCA1 function and associated mechanisms remain unclear. Methods and Results ABCA1 was identified as a potential target of miR-144-3p, based on the results of bioinformatic analysis and the luciferase reporter assay, and downregulated after transfection of cells with miR-144-3p mimics, as observed with real-time PCR and western blot. Moreover, miR-144-3p mimics (agomir) enhanced the expression of inflammatory factors, including IL-1β, IL-6 and TNF-α, in vivo and in vitro, inhibited cholesterol efflux in THP-1 macrophage-derived foam cells, decreased HDL-C circulation and impaired RCT in vivo, resulting in accelerated pathological progression of atherosclerosis in apoE−/− mice. Clinical studies additionally revealed a positive correlation of circulating miR-144-3p with serum CK, CK-MB, LDH and AST in subjects with AMI. Conclusions Our findings clearly indicate that miR-144-3p is essential for the regulation of cholesterol homeostasis and inflammatory reactions, supporting its utility as a potential therapeutic target of atherosclerosis and a promising diagnostic biomarker of AMI.

AIMSAtherosclerosis is a chronic inflammatory disease and represents the major cause of cardiovascular morbidity and mortality. There is evidence that dihydrocapsaicin (DHC) can exert multiple pharmacological and physiological effects. Here, we explored the effect of DHC in atherosclerotic plaque progression in apoE(-/-) mice fed a high-fat/high-cholesterol diet.METHODS AND RESULTSapoE(-/-) mice were randomly divided into two groups and fed a high-fat/high-cholesterol diet with or without DHC for 12 weeks. We demonstrated that cellular cholesterol content was significantly decreased while apoA1-mediated cholesterol efflux was significantly increased following treatment with DHC in THP-1 macrophage-derived foam cells. We also observed that plasma levels of TG, LDL-C, VLDL-C, IL-1β, IL-6, TNF-α and CRP were markedly decreased while plasma levels of apoA1 and HDL-C were significantly increased, and consistent with this, atherosclerotic lesion development was significantly inhibited by DHC treatment of apoE(-/-) mice fed a high-fat/high-cholesterol diet. Moreover, treatment with both LXRα siRNA and PPARγ siRNA made the up-regulation of DHC on ABCA1, ABCG1, ABCG5, SR-B1, NPC1, CD36, LDLR, HMGCR, apoA1 and apoE expression notably abolished while made the down-regulation of DHC on SRA1 expression markedly compensated. And treatment with PPARγ siRNA made the DHC-induced up-regulation of LXRα expression notably abolished while treatment with LXRα siRNA had no effect on DHC-induced PPARγ expression.CONCLUSIONThese observations provide direct evidence that DHC can significantly decrease atherosclerotic plaque formation involving in a PPARγ/LXRα pathway and thus DHC may represent a promising candidate for a therapeutic agent for the treatment or prevention of atherosclerosis.

To evaluate the diagnostic value of serum procalcitonin (PCT) and C-reactive protein (CRP) in patients with acute exacerbation of chronic bronchitis.The PCT and CRP levels were detected in 56 patients with acute exacerbation of chronic bronchitis and 58 healthy control subjects.The serum PCT level and positivity rate were significantly higher in the case group than in the control group (P0.05); the serum CRP level was also significantly higher in the case group (P0.05), but the positivity rate of CRP was similar between the two groups (P0.05). The sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy rate of PCT were higher than those of CRP.Serum PCT is more sensitive than CRP in the diagnoses of acute exacerbation of chronic bronchitis, and can be used as a specific indicator for monitoring the condition of the patients.

To investigate distribution, drug resistance and risk factors of pathogens isolated from septicemic patients in a hospital in the past 6 years.Most of the bacterial isolates were identified with BD Phoenix, and a few isolates were identified manually and with K-B method. Candida isolates were identified with color display plates and K-B method. WHONET5.4 software was used for analysis.The common bacteria isolated form the blood included E. coli, P. aeruginosa, K. pneumoniae, and S. aureu. The gram-negative bacillus from the blood exhibited relatively low resistance to such antibiotics as cefoperazone/sulbactam, imipenem, amikacin, piperacillin/tazobactam, and ceftazidime, and the incidences of E.coli and K. pneumoniae isolates producing extended spectrum beta-lactamase (ESBLs) ranged between 33.3% and 34.9% and between 32.9% and 36.0%, respectively. The gram-positive coccus from blood showed a sensitivity rate of 100.0% to vancomycin and low resistant rates to amikacin and chloramphenicol; the methicillin-resistant rates of S. aureu and coagulase-negative staphylococcus were 26.9%-35.5% and 72.7%-74.3%, respectively. The risk factors of septicemia included hospital stay for over 5 days, venous catheterization, surgeries, puncture, oxygen therapy, urine tract catheterization, and chemotherapy.Blood culture can be of importance in patients with septicemia, and the use of antibiotics should be carefully weighed according to the results of bacterial culture and sensitivity tests of the pathogens isolated from the blood.

To discover the inductive proliferation effect of the RNAi RelB dendritic cells (DCs) on the heterogenic CD4(+)CD25(+) T cells in mice.The proportion of CD4(+)CD25(+) T cells were quantification through flow cytometer when each group of DCs interact with heterogenic T cells in mixed lymphocyte reaction, including control DCs, LPS stimulated DCs, RNAi RelB DCs and LPS stimulated RNAi RelB DCs.Control DCs stimulate more CD4(+)CD25(+) T cells to proliferate than LPS stimulated DCs(P0.01). RNAi RelB DCs and LPS stimulated RNAi RelB DCs significantly improved CD4(+)CD25(+) T cells proliferation compared with LPS stimulated DCs(P0.01). RNAi RelB DCs can also induced CD4(+)CD25(+) T cells to proliferate more than Control DCs (P0.05).RNAi RelB DCs possess powerful potential to induce CD4(+)CD25(+) T cells proliferation, one of characteristics of immature dendritic cells. This type of RNAi RelB DCs can be used as tolerogenic DCs to regulate autoimmunity and other immune responses.

To construct the recombinant lentivirus RNAi vector, and to determine whether the lentivirus mediated short hairpin RNA (shRNA) can inhibit the tissue factor (TF) expression in endothelial cells.Two short hairpin RNAs targeting to human TF were cloned into pENTRTM/U6 plasmid to obtain an entry clone, and the positive clones were verified by sequencing. A recombination reaction was performed between the pENTR/U6 entry construction and pLenti6/BLOCKiTTM-DEST vector, and then the positive clones were confirmed by sequencing. The 293FT cell line was transfected by the above recombined plasmid and lentivirus packing materials, the culture supernatant was harvested, and the virus titer was determined. RT-PCR and ELISA were used to observe the inhibition of TF gene expression after the lentivirus transduction in human umbilical vein endothelial cells.The shRNA sequences targeting to human TF were cloned into the vectors, and an entry clone and an expression clone were constructed successfully, which were proved by sequence determination. Viral particles were packaged in the 293FT cell line, all virus stocks were collected, and the transfection titer was 5*10(5)/transduced unit. RT-PCR and enzyme linked immunosorbent assay demonstrated that the lentivirus stocks could suppress the TF expression in endothelial cells remarkably.Lentivirus RNAi vectors containing human TF gene are successfully constructed, and lentivirus mediated shRNA can inhibit the TF expression in endothelial cells, which may provide a highly effective method for the prevention and treatment of thrombo-embolic diseases.

To detect the changes in serum N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in patients with cardiovascular diseases and explore its clinical significance.Serum NT-proBNP concentrations were measured by electrochemiluminescent immunoassay (ECLIA) in 460 patients with cardiovascular diseases and in 50 normal controls, and echocardiographic examination was performed to determine the left ventricular ejection function (LVEF). Analysis of NT-proBNP was performed for its correlation to New York Heart Association (NYHA) functional classifications LVEF, and risk factors of cardiovascular diseases.The serum LgNT-proBNP concentrations was 3.74 in patients with cardiovascular diseases, significantly higher than that of normal controls (1.42, P0.001). NT-proBNP concentrations also varied significantly among patients with different cardiovascular diseases as shown by one-way ANOVA analysis (F=17.761, P0.001). The NT-proBNP levels increased with the severity of heart failure according to NYHA functional classifications (P0.001), and varied significantly in patients suffering different cardiovascular diseases with the same NYHA functional class. Multivariable regression analysis indicated there were significant correlations of NT-proBNP levels with the patients' age (r=0.152, P0.001), NYHA functional classifications (r=0.725, P0.001), LVEF (r=-0.634, P0.001), and clinica outcomes (r=-0.581, P0.001). Logistic regression analysis identified NT-proBNP level as a strong indicator for cardiovascular events (HR=2.763, P0.01) with close correlation to the treatment results.Serum NT-proBNP level varies significantly with the severity of heart failure and can be indicative of the patients' cardiac function in close correlation to the clinical prognosis, but its value for diagnostic stratification of cardiovascular diseases awaits further investigation.

To explore the relationship between the expression of the nuclear factor-kappaB transcription factor RelB gene and the surface molecules in DC2.4 cell line.DC2.4 cell line was cultured in complete RPMI 1640 medium, whose morphology was observed with optical microscope and the intracellular structures with transmission electron microscope. Flow cytometry was performed to analyze the surface markers of the cells, including MHC-II, CD86 and CD40, and RelB mRNA expression was detected by RT-PCR.Under optical microscope, the cells appeared irregular in shape with obvious dendritic cell processes, and electron microscopy revealed homogenous fat drops and phagocytic vesicles in the cytoplasm. Flow cytometry demonstrated low expression levels of MHC-II and CD40, but high level of CD86 molecules. Low-level expression of RelB mRNA was detected by RT-PCR, resembling its expression level in bone marrow-derived DC with immature phenotype.DC2.4 is a mouse bone marrow dendritic cell line with strong phagocytic capacity, and the low expression of both RelB gene and surface CD40 molecules suggests an immature dendritic cell line.

To explore the expression of avian reticuloendotheliosis viral(v-rel) oncogene-related B (RelB) mRNA in vitro in murine mature and immature myeloid dendritic cells (DCs).The bone marrow was collected from the femur and tibias of C57BL/6 mice in sterile condition. Bone marrow precursors were cultured with recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and interleukin-4 (rmIL-4) to produce immature DCs. Immature DCs were stimulated by LPS 18 h before the end of culture to become mature. DCs' phenotype was detected by flow cytometry (FCM). RelB expression and protein level were determined by RT-PCR and immunofluorescence staining.The expression level of co-stimulatory molecules (CD86 and CD40) and MHC-II class molecule were low in immature DCs, whereas high in mature DCs. RelB expression was significantly higher in mature DCs than in immature DCs (P0.01).RelB expression is closely associated with the maturative situation of DCs. Inhibition of RelB expression in DCs may induce tolerogenic DCs.

To construct small interfering RNA (siRNA) expression cassette targeting murine RelB gene and identify the most effective siRNA sequence against RelB gene in murine bone marrow-derived dendritic cells (DCs).Three expression cassettes namely R1/siRNA, R2/siRNA and R3/siRNA targeting the sites 1027, 302 and 1121 of RelB gene, respectively, were constructed by PCR approach and transfected into cultured murine myeloid DCs by catione liposome Advant-Gene. After incubation for 24 hours in a incubator containing 5% CO(2) at 37 degrees C, the DCs were stimulated by lipopolysaccharide (LPS), and RelB gene expression in DCs were then detected by RT-PCR and immunofluorescence.RT-PCR and immunofluorescence assay showed that the expression of RelB gene in DCs transfected with R2/siRNA could not be upregulated by LPS stimulation, but transfection with R1/siRNA or R3/siRNA failed to produce such effect.R2/siRNA is an effective sequence for RelB silencing, and can be a useful means to construct new tolerogenic DC, RNAi RelB DC, for clinical immunotolerance induction.

To estimate the reliability of heart-type fatty acid-binding protein (H-FABP) for identifying acute coronary syndrome (ACS) in the early stage of chest pain onset.This investigation was conducted based on a small population consisting of 40 healthy individuals, 19 established AMI patients and 20 unstable angina pectoris (UAP) patients. Serum H-FABP concentrations were measured in these subjects by sandwich ELISA, and receiver operating characteristics (ROC) curves for H-FABP for diagnosing AMI and UAP against normal subjects were then generated respectively. The areas under curve (AUCs) were calculated, and 0.5 was defined as the critical value of AUC to evaluate the diagnostic ability.The concentrations of H-FABP in healthy individuals, AMI patients and UAP patients were 1.29+/-0.64, 24.45+/-32.40 and 1.95+/-3.11 ng/ml, respectively; AUC (AMI) and AUC UAP were 0.978 (95%CI: 0.948-1.000) and 0.503 (95% CI: 0.334-0.671) respectively, and the former was significantly greater than 0.5.In the early stage of chest pain onset H-FABP detection is sufficient in distinguishing AMI patients from healthy individuals, but not capable of distinguishing UAP patients from healthy individuals. H-FABP may be used as a diagnostic biochemical marker in the early stage of AMI.

To establish a simple and economic method for in vitro culture of tolerogenic dendritic cells (Tol-DC) from mouse bone marrow to facilitate further research of the application of Tol-DC in autoimmune disease.The dendritic cells were derived from in vitro culture of the precursor cells from mouse bone marrow with recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and interleukin (IL)-4, respectively. Eighteen hours before completion of cell culture, lipopolysaccharide (LPS) was added in the medium for stimulating rmGM-CSF-treated cells, but not IL-4-treated cells. The cells were collected on day 6, and the cell morphology, phenotype and immunofunction were analyzed.DC cultured in vitro had a CD11c(+) expression rate of 76.67% with typical morphology. The DCs without LPS treatment were characterized by moderate expression of class II major histocompatibility complex (MHCII) and low expression of costimulatory molecules, while those with LPS treatment had high expressions of MHCIIand costimulatory molecules.Immature tolerogenic DCs can be derived in large quantity using this method with the potential for use in the treatment of autoimmune diseases.

To evaluate the specificity and sensitivity of immunodiagnosis with the antigen with molecular mass of 32 000 of Angiostrongyliasis cantonensis(AC32).The major antigenic protein AC32 was purified from the antigens of A.cantonensis adult worm by electroelution from SDS-polyacrylamide gels. AC32-enzyme linked immunosorbent assay (ELISA) and adult worm antigen (AWA)-ELISA were used to detect the specific IgG in the sera of normal rats (n=5) and rats with angiostrongyliasis (n=61), sera of healthy individuals (n=50) and patients (n=50) with angiostrongyliasis, schistosomiasis and other parasitosis.The positivity rate in AC32-ELISA of the sera from rats with angiostrongyliasis was 100%; without false positive results or cross reaction between AC32 and the sera of the normal control and patients infected with parasites other than A.cantonensis.AC32 of A.cantonensis is a valuable candidate antigen for immunodiagnosis of angiostrongyliasis with high sensitivity and specificity.

To prepare time-resolved fluoroimmunoassay (TRFIA) kit for detecting human alpha-fetoprotein (hAFP).Sandwich TRFIA kit for hAFP detection was developed using monoclonal antibody of anti-AFP.AFP-TRFIA kit was capable of detecting AFP within the range of 1-1 000 U/ml with sensitivity of 0.17 U/ml and without cross-reactivity with CEA, CA12-5, CA15-3, or CA19-9. The intra- and inter-assay coefficients of variation were (3.3-5.9)% and (3.7-6.5)%, respectively. The prepared AFP-TRFIA reagent could withstand preservation at 4 degrees celsius; for 1 year and at 37 degrees celsius; for 7 days with the cutoff value of 12 U/ml in healthy subjects (n=426). The correlation coefficient of the detection results between this kit and commercially available AFP kit (Wallac OY, Finland) for 60 blood samples was 0.995.The prepared TRFIA kit for hAFP detection meets the demand of clinical application with good sensitivity, precision, specificity, stability and accuracy.

To assess the prognostic value of changes of peripheral blood T cell subsets after thermoradiotherapy for nasopharyngeal carcinoma (NPC).Peripheral blood T cell subsets in 20 normal subjects (control group), 30 NPC patients undergoing thermoradiotherapy, and 20 NPC patients undergoing radiotherapy were detected by flow cytometry.The percentages of CD3+CD4+ and CD8+CD28+ cells were decreased and the percentages of CD3+CD8+ and CD8+CD28- cells increased as compared with the measurements in normal persons. One month after thermoradiotherapy, the percentages of CD3+CD4+ and CD8+CD28+ cells further decreased and the percentages of CD3+CD8+ and CD8+CD28- cells further increased, which continued to worsen 3 months after the treatment and appeared to be related to the survival of the patients.T cell subsets of NPC patients are abnormal and their immune functions depressed in NPC patients within a long period after thermoradiotherapy. CD8+CD28+ and CD8+CD28- T cell subsets can be significant for prognostic assessment in these patients after thermoradiotherapy.

To investigate the distribution and antibiotic sensitivity of the bacteria in the seawater of Xisha in the South China Sea.The samples of the seawater in Xisha were collected for bacterial counting, identification and antibiotic sensitivity tests.Of the 84 bacterial strains isolated from the 80 seawater samples, 38 (45.24%) belonged to Vibrio alginolyticus, 11 (13.10%) to Sphingomonas, 9 (10.72%) to Comamonas terrigena, 7 (8.33%) to Werepseudomonas, 6 (7.14%) to Moraxella, 5 (5.95%) to Cedeceadavisae, and 8 to other bacteria including 2 Aeromona, 2 Acinetobacter, 1 Pantoea agglomerans, 1 Kingella kingae, 1 Shewanella putrefaciens and 1 Suttonella indologenes. Tests of the antibiotic sensitivity showed that all the bacteria were high sensitive to 16 kinds of the antibiotics with the exception of Vibrio alginolyticus, Sphingomonas and comamonas terrigena that were less sensitive to aztreonam or piperacillin and Sphingomonas to cefotaxime or cefepime.The investigation will be of great significance for early-stage prevention and cure of bacterial infection acquired from the seawater.

Background: Although early successes have been achieved in human hand transplant, the changes of immunological parameters in the recipients and their relations to clinical events were not yet known. Methods: In two patients undergoing hand transplantation, we prospectively determined lymphocyte subsets using flow cytometry as well as the serum levels of interleukin (IL)-2, IL-10, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ using enzyme linked immunosorbent assays during the first 6 months after transplantation. Results: The decreases in CD + 3, CD + 4, CD + 8 T cell, the activated T cell (CD + 3/ CD + 25, CD + 3/HLA-DR + ) as well as IL-2, IFN-γ and corresponding significant peak in IL-10 in human hand transplant during the first post-transplant week were observed. Then these parameters recovered to the pre-transplant level except for an even higher level of CD + 8 T cell. The low CD + 4/CD + 8 ratio was been maintained constantly. After 7 wk, IL-2, IFN-γ, and IL-10 decreased to be maintained at a low or undetectable level except for slight increase in IL-10 at post-transplant month 5. There are no significant variation in TNF-a early after transplant. After 3 months, IL-10 was not detected again. Conclusion: The immunosuppressive agents had significantly effects on the immunological status in human hand transplant recipients. These profiles of immunological parameters would be useful data for the future immuno-monitoring in human hand transplant recipients.

To investigate the significance of T lymphocyte subset determination during antithymocyte globulin (ATG) induction therapy in reducing the total drug dose, incidence of complications and cost of treatment in hand allograft.The changes in peripheral blood T lymphocyte subsets (CD3+, CD4+, CD8+, and CD28) were determined by flow cytometry in 3 cases of hand allograft who received ATG treatment.Flow cytometry showed that the percentages of CD3+, CD4+, and CD8+ T lymphocytes, along with the ratio of CD4/CD8, decreased significantly during ATG induction therapy, and the results were consistent in the 3 cases. Long-term continuous changes of peripheral blood lymphocytes were observed after antithymocyte globulin induction therapy.The understanding of the immunological state of the patient with hand allograft after ATG induction therapy by monitoring T lymphocyte subsets may allow adjustment of the total dose of the drugs administered and help prevent the occurrence of complications.

To explore the relationship between serum immunoglobulin (Ig) levels and the positivity rate for auto- antibodies in patients with rheumatoid arthritis (RA).The levels of immunoglobulins (IgG, IgM, IgA) and rheumatoid factor (RF) were measured using Olympus AU600 system, and indirect immunofluorescence and immunoblotting methods were employed respectively for detecting the autoantibodies in the serum from 40 RA patients and 30 normal adults. Statistical analysis of the relation between the Ig levels and autoantibodies was performed.Compared with normal control group, significantly higher serum IgA, IgG and IgM levels were detected in RA patients (P0.01), and at least one autoantibody was present in the serum sample from 26 out of the 40 RA patients, who had the highest RF positivity rate of 20% (8 cases). The serum IgA level in autoantibody-positive patients was significantly higher than that in autoantibody-negative ones (P0.01), and in the 18 patients with serum IgA level beyond the normal range, 15 (83.3%) were positive for autoantibody, while in the 22 patients with normal serum IgA level, the autoantibody positivity rate was only 50% (11/22), showing significant difference (P0.05).Serum immunoglobulins A (IgA) level is closely associated with the positivity rate for autoantibodies in RA patients, and RA patients with serum IgA level beyond the normal range was more likely to be positive for the autoantibodies.

To study the expression of the costimulatory molecule CD28 on peripheral blood CD8(+)T cells from patients with hepatocellular carcinoma (HCC).The expression of CD28 on CD8(+), CD8(+)CD28(-)- and CD8(+)CD28(+) cell subsets in the peripheral blood obtained from 20 HCC patients and 15 healthy controls were assayed by flow cytometry.The elevated percentages of CD8(+) cells, decreased percentages of CD8(+)CD28(+) cells and increased percentages of CD8(+)CD28(-)- cells were observed in the peripheral blood of HCC patients as compared with those of the control group (P0.01). Positive correlation between the percentage of CD8(+)CD28(-)- cells and that of CD8(+) cells were found.CD28 expression on CD8(+) T cells was aberrant in the peripheral blood of HCC patients, and the elevation of CD8(+)CD28(-)- cell number results in the increase of CD8(+) cell number.

To investigate the effects of the dosage of anticancer agents during transcatheter arterial chemoembolization (TACE) on the T cell subsets in patients with hepatocellular carcinoma (HCC).Thirty-six patients with unresectable HCC were randomly divided into 2 groups to receive superselective TACE. Patients in group A (n=18) received low-dose (2-4 mg) mitomycin C (MMC) as the anticancer drug when the tumor was less than 5 cm in diameter; when the tumor ranged from 5 and 8 cm in diameter, 4-6 mg MMC along with 10 mg epirubicin (EPI) was given, and in cases of even larger tumors, 6-8 mg MMC, 10 mg EPI and 100 mg CBP were prescribed. Conventional chemotherapy regimen constituted by 10 mg MMC, 40 mg PI and 300 mg CBP was adopted in group B (n=18). The peripheral blood T cell subsets including CD3(+), CD4(+), CD8(+), NK, CD4(+)/CD8(+) ratio, CD4(+)CD45(+), CD4(+)CD29(+), CD8(+)CD28(+) and CD8(+)CD28- were measured by flow cytometry in both groups before and one week after treatment.The T cell subsets were comparable in the 2 groups before the treatment. After TACE, no significant changes occurred in CD3(+), CD4(+), CD8(+), NK, CD4(+)/CD8(+), CD4(+)CD29(+) or CD8(+)CD28- cells in group A, while significant decrease in CD4(+)CD45(+) and increase in CD8(+)CD28(+) cells were observed (P0.05 and P0.001, respectively). In group B, CD4(+) and CD4(+)CD29(+) levels, together with CD4(+)/CD8(+) ratio, were significantly lower than those before treatment (P0.05), but CD8(+) and CD8(+)CD28- subsets were significantly higher (P0.05).The cellular immune function of HCC patients is significantly impaired by anticancer drugs for TACE at conventional dose, while low-dose of the drugs may enhance the cellular immune function.

Legionella pneumonia is mainly in community-acquired and nosocomial pneumonias caused by the L. pneumophila. The paper reported a case of Legionella pneumonia caused by Legionella pneumophila Sg1 in a man with malignant melanoma. The method for diagnosing Legionella pneumonia by standard culture method，serotyping，PCR-enzymatic digestion analysis and gene sequencing was elaborate. To confirm the diagnosis result of this rapid diagnostic method, sequencing of the bacteria in patient’s sputum partial gene was also carried out. The diagnosis result of this rapid diagnostic method was consistent with the culture method which indicated that it was effective in diagnosing L. pneumophila infection.