Search

Objective: To find the possible targets for the study and treatment of triple-negative breast cancer (TNBC), and to analyze and predict the key genes affecting the prognosis of TNBC by bioinformatics. Methods: Raw data on transcriptome sequencing of clinical specimens from patients with TNBC were searched by searching GEO Datasets in the National Center for Biotechnology Information (NCBI) database. The differential gene was then submitted to the Enrichr website for pathway enrichment. Survival analysis was used to finally identify the most significant differences in the prognosis of patients with TNBC. Results: Only ADAM9 gene showed a significant correlation with the poor prognosis of patients with TNBC ( P< 0.05), and ADAM9 only showed specificity associated with prognosis in patients with TNBC, and was not with other breast cancer types. Conclusion: ADAM9 gene has been proved to be related to the poor prognosis in patients with TNBC. Therefore, ADAM9 gene can be regarded as a possible key gene leading to lymph node metastasis and poor prognosis in patients with TNBC.

Objective: To investigate the features of B7-H1 expression on peripheral myeloid dendritic cells (mDCs) in patients with HIV infection and evaluate the correlations between B7-H1 expression and disease progression., Methods: Peripheral blood samples from 82 treatment-naïve patients with HIV infection, 28 viral complete responders (CRs) under antiretroviral therapy (ART) and 14 healthy controls (HCs) were collected. Flow cytometry was applied to investigate the expression of B7-H1 on mDCs and CD4 cell counts. Plasma HIV-1 viral load was detected by bDNA., Results: The frequency of B7-H1 expression on mDCs were 14.15% +/- 2.63%, 3.31% +/- 0.51% and 0.52% +/- 0.10% in AIDS patients, asymptomatic HIV infected individuals and HCs respectively. As compared with HCs, B7-H1 was significantly up-regulated on mDCs in HIV/AIDS patients. The order was as follows: AIDS patients > asymptomatic HIV infected individuals > HCs (all P < 0.05). Interestingly, the expression of B7-H1 on mDCs in long-term nonprogressors (LTNPs) was 3.12% +/- 1.14%. And it was lower than that in typical progressors (TPs) [8.12% +/- 1.37% (P = 0.001)]. Moreover, the expression of B7-H1 was negatively correlated with CD4 cell counts and positively correlated with plasma viral load in these patients (r = -0.631, P < 0.01 and r = 0.482, P < 0.01 respectively). The expression of B7-H1 on mDCs was significantly lower in ART complete responders than that in AIDS patients (6.59% +/- 1.43% vs 14.15% +/- 2.63%) (P < 0.01). Expression of B7-H1 on mDCs decreased markedly in patients whose CD4 cell counts greatly elevated after a successful antiretroviral treatment., Conclusion: The expression of B7-H1 on mDCs is significantly up-regulated in HIV/AIDS patients. With a close correlation with disease status, it acts as a marker of disease progression.

Objective: Programmed death-1 (PD-1) up-regulation can impair virus-specific CD8(+) T-cell responses during chronic viral infection. Whether PD-1 affects virus-specific CD8 T cells in humans with acute hepatitis B virus (HBV) infection remains unknown. This study aimed to characterize the PD-1 expression during acute hepatitis B (AHB), and further addressed how PD-1 regulates the apoptosis of CD8(+) T cells in this disease., Methods: PD-1 expression on HBV env-183-191-, env-335-343- and CMV pp65-specific CD8(+) T cells were quantitatively analyzed by flow cytometry in peripheral blood from 15 HLA-A2-positive AHB patients. The spontaneous apoptosis indicated by annexin V expression was performed for analyses of the impact of PD-1 expression. We also confirmed the effects of PD-1/PD-L1 pathway on the in vitro apoptosis of CD8(+) T cells through PD-1 blockade assay., Results: PD-1 expression on HBV-specific CD8(+) T cells was significantly up-regulated compared with CMV-specific CD8(+) T cells in the early phase of acute HBV infection. High levels of PD-1 expression significantly correlated with the apoptosis of HBV-specific CD8(+) T cells. Blockade of PD-1 ligation using anti-PD-L1 rescued the HepG2.215-induced CD8(+) T cell apoptosis., Conclusion: PD-1/PD-L1 inhibitory pathway is actively involved in the apoptotic regulation of HBV-specific CD8(+) T cells at early phase of acute HBV infection, and high PD-1 expression by HBV-specific CD8(+) T cells appear to efficiently temper liver damage.

Objective: To investigate the frequencies of immunologically competent cells (ICCs) in the liver-infiltrating lymphocytes (LILs) and peripheral blood and their possible role in pathogenesis in patients with chronic severe hepatitis B (CSHB)., Methods: LILs were isolated from the liver tissue samples from 11 CSHB patients and 5 normal controls (NCs) by the method of combined grinding with semi-frosted microscopic slides and sedimentation of hepatic cells. The frequency of isolated ICCs, including CD3(+), CD4(+), and CD8(+) T-cells, NK cells, NKT cells, and B cells was examined and compared with that of the circulating ICCs in the CSHB patients. Comparison was conducted between the CSHB patients and the controls., Results: (1) In the CSHB patients, the frequencies of CD4(+) T cells and B cells in LILs were 17% +/- 6% and 3.0% +/- 1.0% respectively, both significantly lower than those in the circulating blood (32% +/- 8% and 21.4% +/- 12.2% respectively, both P < 0.01); however, the frequencies of CD8(+) T cells, NK cells, and NKT cells in LILs were 38% +/- 13%, 34% +/- 18%, and 10% +/- 4% respectively, all significantly lower than those in the circulating blood (26% +/- 6%, 15% +/- 9%, and 6% +/- 4%, all P < 0.05). (2) The frequencies of infiltrating CD3(+) T cells and CD4(+) T cells of the CSHB patients were both significantly higher than those of the NCs (P = 0.042 and P = 0.001); and the frequency of infiltrating CD8(+) T cells of the CSHB patients was higher than that of the NCs, and the and the frequencies of infiltrating NK cells and NKT cells in LILs were lower than those of the NCs, however, not significantly. (3) Compared with the liver tissues from the NCs, the liver tissues from the CSHB patients exhibited a significantly higher ratio of liver-infiltrating CD4(+) T cells to peripheral blood CD4(+) T-cell subsets (P = 0.001), and significantly lower ratios of liver-infiltrating NKT cells and B cells to the peripheral blood NKT-cells and B cells (P = 0.029 and P = 0.001 respectively)., Conclusion: The abundant infiltrating immune active cells, especially the CD4(+) T cells, CD8(+) T cells, and NK cells, may be the causal factors that drive the progressive development of CSHB.

Objective: CD4(+)CD25(high) regulatory T cells (Treg) have been shown to play an important role in maintaining peripheral tolerance against self and foreign antigens, and in suppressing T cell immune response. Our aim was to characterize circulating Treg in HBV-infected patients., Methods: Treg in peripheral blood from 72 chronic hepatitis B (CHB) patients, 16 acute hepatitis B (AHB) patients and 32 healthy subjects were quantitatively analyzed using flow cytometry. Serum HBV markers were evaluated for each subject. HBV-DNA levels were measured using real-time RT-PCR., Results: CHB patients presented a higher fraction of circulating Treg (3.9% +/- 1.4%) than those in AHB patients (3.1% +/- 0.9%) (P < 0.05), but were similar to healthy controls (3.5% +/- 0.7%). CHB patients with greater than 10(7) copies/ml of serum HBV DNA loads had a higher frequency (4.5% +/- 1.9%) of circulating Treg than health controls (P < 0.01) and the patients with less than 10(7) copies/ml of serum viral loads (3.4% +/- 0.7%). A correlation was found between circulating Treg and HBV DNA level (r = 0.32, P < 0.01). Furthermore, Treg was more frequent in convalescent phase (6.0% +/- 1.7%) than in early acute phase (3.0% +/- 0.6%)., Conclusion: Increased peripheral Treg is found to be associated with HBV replication in chronic hepatitis B. In acute HBV infection, Treg is less frequent in early phase. The related mechanisms is under further investigation.

Objective: To investigate pathophysiological changes of the remnant esophagus and gastric cardia in patients who underwent esophagectomy for cancer, and to provide objective evidences for the improvement of the postoperative quality of life., Methods: The function of the remnant esophagus and intrathoracic stomach in patients who underwent esophagectomy for cancer were assessed objectively. The methods that we used were gastric scintigraphy, esophageal manometry, 24-hour esophageal pH monitoring, electronic gastroscopy, videofluoroscopy, and DeMeester scoring system for the assessment of heartburn. Findings were recorded and compared with normal controls., Results: After esophagectomy for cancer, the emptying of intrathoracic stomach was delayed (t = 7.105, P < 0.01) and improved over time, but could not reach normal one year after surgery (t = 2.9, P = 0.016). In patients who had undergone esophagectomy for cancer, the contracting pressure of the upper esophageal sphincter and resting pressure of the remnant esophagus were higher than that in normal controls (t = 2.275, P = 0.03; t = 2.16, P = 0.039 respectively). 89.7% of patients who had undergone esophagectomy had gastroesophageal reflux measured with 24-hour pH monitoring. The extent of reflux was less severe when patients were in a semi-reclining position than in a prostration position (t = 3.074, P = 0.005)., Conclusion: After esophagectomy for cancer, delayed emptying of the intrathoracic stomach is improved gradually over time, but it is inaccessible to normal level. Gastroesophageal reflux extensively exists in patients who underwent esophagectomy for cancer, but it can be lessened by taking semi-reclining position.

Objective: To investigate the changes of circulating dendritic cell (DC) sunsets in chronic hepatitis B patients treated with alpha-interferon and correlation thereof with therapeutic effectiveness., Methods: Twenty-five patients with chronic hepatitis B. 24 males and 1 females, aged 18 approximately 43, were treated with subcutaneous injection of alpha-interferon for 24 weeks and then followed up for another 24 weeks. Blood routine, liver function, renal function, and serum HBV DNA were examined before treatment, and 12, 24, and 48 weeks after beginning of treatment. Peripheral blood samples were collected to detect the DC subsets by flow cytometry., Result: Those patients with complete or partial recovery of alanine transaminase (ALT) and negative seroconversion or decrease of HBV DNA with/without negative seroconversion of HBeAg and HBsAg were grouped as alpha-interferon responders, and those failing to reach the above criteria were grouped as nonresponders. The negative seroconversion rates of HBV DNA and HBeAg, and positive seroconversion rate of anti-HBe were 35%, 40%, and 10 respectively after 24 weeks' treatment and were 40%, 33.3%, and 20% respectively after 48 weeks' treatment. The numbers of myeloid dendritic cells (mDCs) were 16.5 +/- 5.51 x 10(6)/L, 9.86 +/- 5.2 x 10(6)/L, 9.20 +/- 3.19 x 10(6)/L, and 10.0 +/- 3.64 x 10(6)/L respectively in the alpha-interferon responders (P = 0.045); and were 14.41 +/- 6.62 x 10(6)/L, 14.0 +/- 5.27 x 10(6)/L, 10.40 +/- 4.6 x 10(6)/L, and 12.3 +/- 5.23 x 10(6)/L respectively in the non-responders (P = 0.283) before treatment, and after 12, 1 24, and 48 weeks' treatment respectively. The numbers of plasmacytoid dendritic cells (pDCs) were 5.91 +/- 2.35 x 10(6)/L, 4.25 +/- 2.00 x 10(6)/L, 3.30 +/- 1.55 x 10(6)/L, and 4.32 +/- 1.59 x 10(6)/L respectively in the alpha-interferon responders (P = 0.004); and were 5.10 +/- 1.72 x 10(6)/L, 4.06 +/- 1.67 x 10(6)/L, 3.89 +/- 1.25 x 10(6)/L, and 4.06 +/- 8.12 x 10(6)/L respectively in the non-responders before treatment, and after 12, 24, and 48 weeks' treatment respectively ((P = 0.211)., Conclusion: Effective on CHB, alpha-interferon induces the percentages and numbers of circulating DC subsets, especially in the-interferon responders.

Objective: To investigate the dynamic changes of viral loads and immunocytes during the in vitro culture of peripheral blood mononuclear cells (PBMC) from HIV carriers., Methods: The PBMCs from 14 HIV-infected individuals and 6 healthy persons were incubated in serum-free AIM-V medium containing cocktail cytokines. The phenotype of CD3, CD4, CD8, CD3CD56 and CD25 was identified by flow cytometric analysis every two days. The production of cytokines in the supernatants, including IL-1alpha, IL-12, TNF-alpha and IL-10 was measured by ELISA. The supernatant HIV-1 RNA load was also determined by Real-time fluorescent PCR., Results: During a 21-day incubation period, The PBMCs multiplied approximately 60.7-fold and 16.8-fold respectively in the healthy controls and 7 out of the 14 HIV-infected subjects, however failed to multiply in the remaining 7 HIV-infected subjects. The expanded cells were phenotypically shown a heterogeneous cellular population with 23.3%-35% for CD3(+)CD4(+) T cells and 58.7%-72% for CD3(+)CD8(+) T cells, and approximate 17% CD3(+)CD56(+) cells at 16-day incubation for HIV-infected cases. HIV-1-positive PBMCs were found to produce an elevated ratios (value range 6.01 - 48.04) of IL-12:IL-10 compared to healthy individuals (6.65 - 10.2) at 16-day incubation. Furthermore, serial analyses of HIV-1 RNA levels showed an inverted V type dynamic change during 16 day in vitro incubation period., Conclusion: In vitro expansion of functional immunocytes of HIV-1 carrier origin is feasible and may facilitate the autologous antiviral immune therapy for HIV-infected patients.

Objective: To investigate the characteristics of phenotype and function of myeloid dendritic cells (mDCs) pulsed with HBV antigens derived from HBV-associated hepatocellular carcinoma (HCC) patients., Methods: Peripheral blood mononuclear cells (PBMCs) were collected, using blood cell separator, from 21 primary HCC patients and 4 healthy donors. mDCs were propagated in serum-free AIM-V medium in the presence of cytokine cocktail, and pulsed with HBcAg or HBsAg. After 9 days' incubation, the phenotypic patterns of mDC were characterized by flow cytometry and the levels of IL-10 and IL-12 produced by mDCs were analyzed by ELISA. Autologous T cells proliferation stimulated by mDC was tested by non-radioactive cell proliferation assay kit., Results: The expression rates of CD80, CD86, CD40, and HLA-DR in the mDCs pulsed with HBcAg were 55% +/- 26%, 80% +/- 13%, 70% +/- 13% and 73% +/- 24% respectively, significantly higher than those of the control group (29% +/- 25%, 35% +/- 18%, 44% +/- 26% and 45% +/- 23% respectively, all P < 0.05). Among the expression rates of surface molecules of the mDCs pulsed with HBsAg only the expression rate of HLA-DR was significantly higher than that of the un-pulsed mDCs (63% +/- 15% vs 45% +/- 23%, P < 0.05). T cell proliferation assay revealed an impaired allostimulatory capacity of mDCs in HCC and the stimulatory capacity of the mDCs pulsed with HBcAg to induce proliferation of autologous T cells was more powerful than that pulsed with HBsAg (P < 0.05). The levels of IL-10 and IL-12 produced by the mDCs pulsed with HBsAg were (35 pg/ml +/- 9 pg/ml and 135 pg/ml +/- 63 pg/ml respectively, both significantly lower than those pulsed with HBcAg (236 pg/ml +/- 95 pg/ml and 733 pg/ml +/- 212 pg/ml respectively, both P < 0.05)., Conclusion: Pulsation of mDCs in vitro by HBcAg or HBsAg enhance the expression of CD80, CD86, CD40, and HLA-DR, and increase the ability of mDCs to stimulate the proliferation of autologous T lymphocytes.

Objective: To investigate the difference of host immune response specific to hepatitis B virus (HBV) infections between acute self-limited and chronic persistent hepatitis by quantitative analysis of HLA-A * 2402-restricted HBcAg-specific cytotoxic T lymphocyte cells (CTL) cells., Methods: The frequency of HBV-specific CTL cells in the peripheral blood mononuclear cells (PBMCs) from 20 patients infected with HBV were quantified by ELISPOT assays and flow cytometry using one HLA-A * 2402-HBV, 7 with acute HB and 13 with chronic HB, peptide tetrameric complex., Results: High frequencies of circulating HBcAg-specific CTL cells were detected in most individuals with acute HBV infection while the number of these cells was significantly reduced at the convalescent stage. HBcAg-specific CTL cells were not detected in the PBMC from individuals with chronic HBV infection except for one patient with an acute infection exacerbation., Conclusion: HBcAg-specific CTL cells may play a crucial role in complete clearance of HBV from patients with acute HBV hepatitis.

Objective: To investigate the dynamic changes of dendritic cell subsets in peripheral blood of patients infected with severe acute respiratory syndrome (SARS) and evaluate their roles in the immunopathogenesis of SARS., Methods: Flow cytometry was applied to study the dynamic alteration of the number and frequencies in circulating DC cell subsets in 30 SARS patients including critical SARS (n = 11) and general SARS (n = 19). The reasons and clinic significances of the peripheral blood DC subsets changes in SARS patients were also analyzed in our study., Results: The patients in critical status had a 9-week course of disease, longer than the 6-week course observed in subjects in general status. The frequency of peripheral DC cell subsets significantly dropped beginning from the onset of symptom in SARS patients and was maintained at significant low levels during the following 4 - 5 weeks, 1.7 +/- 1.8, 5.3 +/- 5.0/ micro l for DC1, 0.57 +/- 1.02, 0.98 +/- 1.11/ micro l for DC2 for cases in critical and general statuses, respectively, compared with healthy subjects; more importantly, the pDC2 even disappeared in the patients who died from SARS diseases. The possible reasons responsible for the alteration of DC subsets in peripheral blood is likely to be the direct attack of SARS-CoVin circulation and be partially involved the application of large dose of steroid. The frequency in DC cell subsets returned to normal level in convalescent stage., Conclusion: Our results showed SARS patients had a significant decrease of circulating DC cell subset frequency, which maybe lead to the host immunodeficiency response to SARS-associated coronavirus (SARS-CoV).

Objective: To investigate the alteration of the cellular profiles of T lymphocyte subsets and dendritic cell subsets in peripheral blood of primary hepatocellular carcinoma (HCC) patients after being transfused with autologous cytokine-induced killer cells (CIK) in patients, then to evaluate the clinical efficacy of the immune therapeutic strategy., Methods: Peripheral blood mononuclear cells (PBMCs) from 13 patients with primary were collected using blood cell separator, and expanded in the fresh AIM-V medium in the presence of cytokine cocktail including interferon-gamma (IFN-gamma), monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2). The phenotypic patterns of CIK cells were longitudinally characterized by flow cytometry on day 0, 4, 7, 10,13 and 15 during the incubation period. PBMCs obtained from HCC patients before or after CIK cells transfusion into bodies to assay the changes of proportion of DC1 or DC2 in peripheral blood., Results: After in vitro incubation for 14 or 15 days, a large of CD3(+)CD56(+) cells were produced from their progenitors and the percentages of CD3(+)CD8(+), CD3(+)CD56(+), CD25(+) cells significantly increased from 33.5% +/- 10.1%, 7.7% +/- 2.8%, and 12.3% +/- 4.5% at the beginning to 36.6% +/- 9.0% (P < 0.05), 18.9% +/- 6.9% (P < 0.01), and 16.4% +/- 5.9% (P < 0.05) at the day 15, respectively. In contrast, the percentages of CD3(+)CD4(+) and NK cells displayed no significant difference. The percentages of CD3(+), CD3(+)CD8(+) cells was held at a higher level during the whole incubation period, however those of the CD25(+), and CD3(+)CD56(+) cells began decreasing on day 7 and day 13, respectively. The proportion of type I of dendritic cells (DC1) and type II of dendritic cells (DC2) subsets increased from 0.59% +/- 0.23% and 0.26% +/- 0.12% before CIK cell transfusion to 0.85% +/- 0.27% and 0.43% +/- 0.20% (all P < 0.01) after CIK cell transfusion. The symptom of HCC patients receiving the CIK cell therapy was markedly ameliorated, and not side effect was seen in the treatment., Conclusion: Our results indicated that autologous CIK cells is able to boost the cellular immunological function in HCC patients, which probably provide a potent immune therapeutic strategy for HCC patients.

Objective: Investigate the features of outbreak epidemic, clinical disease progression of the first SARS cases in Beijing and evaluate the efficacy of therapeutic regimen., Methods: Twenty-nine patients (11 men and 18 women, 20 - 74 years old age range) were diagnosed with infectious SARS and admitted in our hospital from the March 5th to April 14th, 2003 in this study. The data of clinical presentation and disease progression of all the patients including index subject as the infectious SARS resource patient, her family infected members and 21 health care workers were abstracted., Results: The first SARA outbreak in Beijing was characterized with the cluster feature of resource patient family members and health care providers. The incubation period ranged from 2 to 14 days. All the patients had a fever (temperature > 38 degrees C for over 24 hours) and other manifestations as reported before. Serial chest radiographs showed progressive pathologic air-space disease. Twenty patients showed the severe syndrome with various time ranged from 1 day to 14 days. Two patients died of progressive acute respiratory distress disease. The histologic analysis of one death patient showed diffuse alveolar damage in the two lungs. Twenty-six patients receiving the combined therapy including use of corticosteroid, antiviral ribavirin agents after the onset of symptoms and showed they had an acute self-limited disease course. The oldest patient (74 year old, male) received the healthy convalescent plasma infusion (50 ml) from recovered SARS subject and completely recovered within 21 days, having a shorter disease course., Conclusion: SARS is a kind of new self-limited and acute infectious disease. Early diagnosis, early isolation, early antiviral therapy for patients and efficient prevention for health care providers are urgently recommended. In particular, a combinational therapy of use of antiviral agents, preventive antibacterial antibiotics and pulsed dosage of corticosteroid can efficiently raise the clinical recovery rate and decrease mortality of SARS patients.

Objective: To investigate the number, phenotype, and interferon-alpha (INF-alpha) of type II dendritic cells (DC2) in persons with hepatitis B and evaluate the role of DC2 subset in the immunopathogenesis of chronic HBV infection., Methods: Peripheral blood was extracted from 103 hepatitis B (HB) virus-infected persons, including 11 cases of HB virus (HBV)-infected persons, 11 cases of acute HB, 81 cases of chronic HB, and 11 cases of asymptomatic HBV infection, and 25 healthy blood donors used as controls. Flow cytometry was used to calculate the number and the phenotype of circulating DC2. Ultraviolet-inactivated herpes simplex virus (HSV)-1 was added into the suspension of peripheral blood mononuclear cells (PBMCs) and then co-cultured for 24 hours to stimulate the production of INF-alpha by DC2 that was examined by ELISA assay., Results: The number of DC2 in patients with chronic HB was 3.3 +/- 1.0 10(6)/L, significantly lower than that in the healthy controls (7.2 +/- 2.4 10(6)/L, P < 0.01). However, the number of DC2 was not significantly different between any other groups. The proportion of GS2 to PBMCs in the patients with chronic HB was 1.12 +/- 1.13 approximately 0.22 +/- 0.10, all significantly lower than that in the healthy controls (0.32% +/- 0.13%, P < 0.01). However, the proportion of GS2 to PBMCs was not significantly different between any other groups. The decrease of number of DC2 and that of proportion of DC2 to PBMCs in patients with chronic HB were related with the progress of disease. The INF-alpha concentration in the suspensions of PBMCs of different groups without stimulation by HSV-1 were low and there was no significant difference in INF-alpha concentration between different groups. The INF-alpha concentration in the suspension of PBMCs of healthy controls was 789 +/- 82 pg/ml, significantly higher than those of the patients with acute HB (161 +/- 36 pg/ml) and the patients with chronic HB (183 +/- 113 pg/ml, 147 +/- 39 pg/ml, and 156 +/- 39 pg/ml, all P < 0.05). However, there was no significant difference between the patient groups (all P > 0.05)., Conclusion: The number and INF-alpha producing function of DC2, and the numbers of NK cells and CD8+ T cells in peripheral blood of patients with chronic HB decrease significantly, which results the deficiency of HBV-specific immune response.