Effect of moxibustion on skin lesions and immune inflammatory response in psoriasis mice / 中国针灸

OBJECTIVE@#To observe the effect of moxibustion on skin lesions and immune inflammatory response in psoriasis mice, and to explore the possible mechanism of moxibustion for psoriasis.@*METHODS@#A total of 32 male BALB/c mice were randomly divided into a normal group, a model group, a moxibustion group and a medication group, 8 mice in each group. Psoriasis model was induced by applying 5% imiquimod cream on the back for 7 days in the model group, the moxibustion group and the medication group. At the same time of model establishment, the moxibustion group was treated with suspension moxibustion on skin lesions on the back, 20 min each time, once a day; the medication group was treated with 1 mg/kg methotrexate tablet solution by gavage, once a day. Both groups were intervened for 7 days. The daily changes of skin lesions were observed, and the psoriasis area and severity index (PASI) score was evaluated; the histopathological changes of skin lesions were observed by HE staining; the positive expression of proliferating cell nuclear antigen (PCNA) and T lymphocyte surface marker CD3 were detected by immunohistochemistry; the expression level of serum interleukin (IL) -17A was detected by ELISA, and the relative expressions of tumor necrosis factor-α (TNF-α), IL-1β and IL-6 mRNA in skin lesions were detected by real-time PCR.@*RESULTS@#The increased and hypertrophy scale, dry skin, red and swollen epidermis and obvious infiltration were observed in the model group, and each score and total score of PASI were higher than those in the normal group (P<0.01). The scale score, infiltration score, and total score of PASI in the moxibustion group were lower than those in the model group (P<0.01); the infiltration score and total score of PASI in the medication group were lower than those in the model group (P<0.01, P<0.05). The inflammatory cell infiltration in the model group was obvious, and the thickness of epidermal layer was increased compared with that in the normal group (P<0.01); the inflammatory cell infiltration and Munro micro abscess were decreased in the moxibustion group and the medication group, and the thickness of epidermal layer was decreased compared with that in the model group (P<0.01). Compared with the normal group, the positive cell number of PCNA and T was increased (P<0.01), and the body mass was decreased, and the spleen index was increased (P<0.01), and the expression of serum IL-17A and the relative expression of TNF-α, IL-1β and IL-6 mRNA in the skin lesions was increased in the model group (P<0.01). Compared with the model group, the positive cell number of PCNA and T was reduced (P<0.01), and the spleen index and the relative expression of TNF-α, IL-1β and IL-6 mRNA were reduced (P<0.01) in the moxibustion group and the medication group; the body mass of mice in the moxibustion group was higher than that in the model group (P<0.01); the content of serum IL-17A in the medication group was lower than that in the model group (P<0.01); the relative expression of TNF-α, IL-1β mRNA in the moxibustion group was higher than that in the medication group (P<0.01).@*CONCLUSION@#Moxibustion could effectively improve the scale and infiltration of skin lesions in psoriasis mice. Its mechanism may be related to inhibiting inflammatory response and regulating immunity.

Analysis of Factors Influencing Prognosis of 87 Adults with Ph Chromosome Negative ALL / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the factors influencing total complete remission (CR), recurrence, disease-free survival (DFS) rate and overall survival (OS) rate in adults with Philadelphia (Ph) chromosome negative acute lymphoblastic leukemia (ALL) and the effect of subsequent allogeneic hematopoietic stem cell transplantation (allo-HSCT) on prognosis.</p><p><b>METHODS</b>The clinical data of 87 adult patients with Ph negative ALL were retrospectively analyzed, the CHOP regimen plus L-asparaginase (L-Asp) was used for the induction therapy, and the CHOP+ modified Hyper-CVAD or methotrexate was set up as the consolidation chemotherapy regimen. After consolidation chemotherapy for 3-6 courses, 45 patients (51.72%) received allo-HSCT , and 42 patients (48.28%) continually received the maintained consolidation chemotherapy. The average follow up time of the surviving patients was 40.13 (3-60 months).</p><p><b>RESULTS</b>Out of 87 patients with PhALL one patient died (1.15%). In 86 patients who could be evaluated, 68 cases (79.67%) reached CR at the end of 1 course, 80 cases obtained CR (93.02%). Multivariate regression analysis showed that the enlargement of lever, spleen and lymphomode, WBC count≥ 100×10/L were affecting factors for total CR (P<0.05). Among 80 cases with CR, 27 cases (33.75%) relapsed, 5 years' overall survival (OS) rate and disease-free survival (DFS) rate were 47.50% and 45.00% respectively. Multivariate regression analysis yet showed that the induction chemotherapy without L-Asp, presence of CNS leukemia at diagnosis, absence of allo-HSCT and no CR after indution chemotherapy for 4 weeks were affecting factors for relapse and poor prognosis of patients (P<0.05). According to 4 prognostic factors such as presence of CNS leukemia or no, WBC count≥100×10/L or no, induction chemotherapy with L-Asp or no and CR after induction chemotherapy for 4 weeks or no, 86 patients were divided into low-risk group (without poor prognostic factor), middle-risk group (with 1 poor prognostic factor), high-risk group (with 2-4 poor prognostic factors). Statistical results showed that allo-HSCT treatment in low-risk group had no significant effect on OS and DFS (P>0.05). The rate of OS and DFS in middle and high-risk group were significantly higher than those of patients without allo-HSCT treatment (P<0.05).</p><p><b>CONCLUSION</b>Patients with central nervous system leukemia, high white blood cell count (≥100×10/L), induction chemotherapy without L-Asp, no CR after 4 weeks of chemotherapy and absence of allo-HSCT treatment are the factors influencing the prognosis of adult patients with Ph negative ALL, so the patients with those poor prognostic factors should take active treatment of allo-HSCT.</p>

Renal artery embolization with exploratory laparotomy for management of massive colonic hemorrhage after renal biopsy: a case report / 南方医科大学学报

We report a case of massive colonic hemorrhage after renal biopsy managed by renal artery embolization combined with exploratory laparotomy. Clinicians must be alert for such rare anatomical abnormalities as ectopic colon behind the kidney and the risk of colonic hemorrhage following renal biopsy. In this case, artery embolization combined with exploratory laparotomy successfully and quickly stopped the bleeding and avoided possible organ resection.

Effects of peritoneal dialysis solution on apoptosis and intracellular free calcium, cell surface ICAM-1 expression of rat peritoneal mesothelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of peritoneal dialysis solution (PDS) on apoptosis and intracellular free calcium([Ca(2+)]i), cell surface ICAM-1 expression of rat peritoneal mesothelial cells (RPMCs).</p><p><b>METHODS</b>The RPMCs apoptosis rate were determined by flow cytometry. [Ca(2+)]i in the cells were monitered the fluorescence at 528 nm by confocus laser microscopy. Cell surface ICAM-1 expression were detected by flow cytometry.</p><p><b>RESULT</b>After PDS treatment for 1 h, the RPMCs apoptosis rate were increased. Such increase was more manifest with higher glucose concentration in PDS and longer treatment time of the cells. At the same times, after 3 hours, ICAM-1 expressions of the PDS containing glucose and mannitol are all increased. With the increase of glucose concentrations, the descend of [Ca(2+)]i levels were aggravated.</p><p><b>CONCLUSION</b>PDS containing high- concentration glucose can induce significant apoptosis of RPMCs in vitro. This may be related with the enhanced level of ICAM-1 expressions and the decreased level of [Ca(2+)]i. Which may due to the occurrence of peritoneal fibrosis and ultrafiltrate failure in patients suffering long term peritoneal dialysis.</p>

Comparison of two rat models of IgA nephropathy / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the methods for rapid establishment of rat models of IgA nephropathy.</p><p><b>METHODS</b>Forty female SD rats weighing 160-200 g were randomized into 3 groups. In group A, the rats received intravenous injection of staphylococcal enterotoxin B (SEB) and oral bovine serum albumin (BSA), and in group B, CCl4 was injected subcutaneously in addition to the above treatments; the rats in group C received no treatments to serve as the normal control group. The rats were sacrificed 10 and 14 weeks after the treatment for biochemical testing of the arterial blood and histopathological and IgA immunofluorescence examination of the renal tissues. The twenty-four-hour urine was collected at 10, 12, and 14 weeks after the treatments for detecting the urine proteins.</p><p><b>RESULTS</b>Compared with the control group, the rats in groups A and B showed significantly increased serum creatinine, urine nitrogen and protein levels. Pathological examination of the renal tissue showed mild to moderate mesangial expansion and mesangial cell proliferation in groups A and B, without obvious difference between the two groups; but hematuria and proteinuria occurred earlier in group B with stronger IgA immunofluorescence than in group A.</p><p><b>CONCLUSION</b>Both of the methods used in group A and group B can successfully induce IgA nephropathy in rats, but in group B, hematuria and urineprotein occurs earlier and IgA immunofluorescence is more stronger. Therefore intravenous SEB injection combined with oral BSA and subcutaneous CCl4 administration is a better method for time-efficient establishment of rat models of IgA nephropathy.</p>

Foxp3-transfected CD4+ CD25- T cells suppress function of dendritic cells / 中国实验血液学杂志

To explore the relationship between expression of Foxp3 gene and immune activity of CD4(+) T cells, the Foxp3 gene was transfected with retroviral vector and applied to forcedly express Foxp3 protein in naive CD4(+)CD25(-) T cells, and then the effect of transfected CD4(+)CD25(-) T cells on immune co-stimulatory molecules and immune function of dendritic cells (DCs) was investigated, and the dependence of direct contact between Foxp3-transfected CD4(+)CD25(-) T cells and DCs was clarified by Transwell test. The results showed that through transfection of retroviral vector, CD4(+)CD25(-) T cells model expressing Foxp3 was established. At 1 week after transfection, proportion of T cells expressing Foxp3 was 38%. CD4(+)CD25(-) T cells forcedly expressing Foxp3 could play immune suppression role in vitro and induce down-regulation of CD80 and CD86 expression on the membrane of DCs. The lymphocyte proliferation test in vitro indicated that Foxp3 transfected CD4(+)CD25(-) T cells could inhibit effect of DCs on activation of allo-lymphocytes. It is concluded that the effect of Foxp3-transfected CD4(+)CD25(-) T cells on DC depends on intercellular direct contact.

Induction of tolerogenic dendritic cells by membrane-bound HLA-G in vitro / 中国实验血液学杂志

In order to study how to induce tolerogenic dendritic cells in vitro and its mechanism, the K562 cells transduced with HLA-G construct were used to co-culture with DC. Then their related immunological changes, such as membrane molecules CD80, CD86, ILT3 and ILT4 expression levels were detected by flow cytometry. Allogeneic proliferation of peripheral blood mononuclear cells (PBMNC) was detected by mixed lymphocyte reaction. The results showed that CD80 and CD86 expressions on DC were downregulated, while ILT3 and ILT4 expressions were upregulated after co-culturing with K562-HLA-G cells. The DCs were less able to stimulate the allogenic PBMNC. It is concluded that the membrane-bound HLA-G can upregulate expression of inhibitory receptors ILT3 and ILT4, inducing tolerogenic DC in vitro, which may provide a novel strategy for transplant tolerance induction.

Study of paired immunoglobin-like receptor B expression on dendritic cells and its relationship with immune tolerance in mouse / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of paired immunoglobin-like receptor B (PIR-B) on dendritic cells (DCs) and its relationship with tolerogenic DCs (T-DCs) in mouse.</p><p><b>METHODS</b>DC2.4 cells, an immature dendritic cell line derived from C57BL/6 mouse, were stimulated by lipopolysaccharide (LPS) for 48 h to induce the mature dendritic cells (mDC) and cultured respectively with the recombined mouse interleukin-10 (rmIL-10) or recombined human transforming growth factor beta1 (rhTGF-beta1) to develop the tolerogenic dendritic cells (T-DC). Special small interference RNA (siRNA) molecular of PIR-B was chemically synthesized and transfected into DC2.4 cells (si-DC) by lip2000. The expression of PIR-B on DC2.4 cells was measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot. The allogeneic lymphocyte proliferative capacity of DCs was measured by mixed lymphocyte reaction (MLR) using 3H-thymidine incorporation test. The concentration of IFN-gamma in supernatants of MLR was analyzed by ELISA.</p><p><b>RESULTS</b>Semi-quantitative RT-PCR and Western blot showed that PIR-B mRNA and protein were expressed on DC2.4 cells. RmIL-10 and rhTGF-beta1 induced the higher PIR-B mRNA and protein level on T-DCs (Relative values were 0.51 +/- 0.08 and 0.58 +/- 0.23; 0.85 +/- 0.07 and 0.87 +/- 0.14; 0.79 +/- 0.10 and 0.85 +/- 0.34, respectively) (P < 0.05). LPS down-regulated the PIR-B expression on mDC (0.35 +/- 0.10 and 0.32 +/- 0.04) (P < 0.05). The PIR-B mRNA and protein expression were inhibited by siRNA transfection (decreased by 78.9% and 74.2% respectively after 48 h interference) (P < 0.05). DC2.4 cells stimulated the proliferation of BALB/c mouse allo-genetic spleen cell. The mDC enhanced alloreactivity in MLR and the IFN-gamma secretion in supernatants. The T-DCs alleviated the allo-genetic spleen cell proliferation (P < 0.05) and IFN-gamma secretion in MLR (P < 0.05). Silence of the PIR-B expression (si-DC) also promoted of alloreactivity and enhanced the IFN-gamma secretion in MLR (P < 0.05).</p><p><b>CONCLUSION</b>High expression of immune inhibition receptor PIR-B is one of the general features and molecular mechanism of dendritic cells to acquire immune tolerance in mouse.</p>

Effects of leflunomide combined with hormone therapy for refractory IgA nephropathy / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects and safety of leflunomide combined with hormone therapy for refractory IgA nephropathy.</p><p><b>METHODS</b>Thirteen patients with refractory IgA nephropathy were treated with leflunomide and hormone therapy, and the clinical data were collected and evaluated before and in weeks 2, 4, 8, 12, 16, 20 and 24 during the treatment.</p><p><b>RESULTS</b>Leflunomide therapy significantly improved proteinuria (Plt;0.001) and increased serum albumin in these patients (Plt;0.001) but caused no significant changes in serum creatinine (Pgt;0.05). Only mild tolerable adverse effects were observed.</p><p><b>CONCLUSION</b>Leflunomide combined with hormone therapy can be one of the safety and effective choices for treatment of refractory IgA nephropathy.</p>

Effects of peritoneal dialysis solution on proliferation and apoptosis of rat peritoneal mesothelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of peritoneal dialysis solution (PDS) on proliferation and apoptosis of rat peritoneal mesothelial cells (RPMCs).</p><p><b>METHODS</b>The proliferation of RPMCs treated with PDS containing glucose of different concentrations for different times in vitro were examined by MTT colorimeric assay. The cell cycles and cell apoptosis rate were determined by flow cytometry.</p><p><b>RESULTS</b>After PDS treatment for 1 h, the cell proliferation inhibition rate, percentage of cells at G(0)/G(1) stage and early cell apoptosis rate increased, and such increment was more manifest with higher glucose concentration in PDS and longer treatment time of the cells. After treatment with PDS containing 4.25% glucose for 3 h, the proliferation inhibition rate of the RPMCs reached 44.12%, the percentage of cells at the G(0)/G(1) stage amounted to 71.95% and the early apoptosis rate reached 23.59%, which was several times higher than that of the negative control and 1.5% glucose/PDS groups, and also higher than that of 4.25% mannitol/PDS groups.</p><p><b>CONCLUSION</b>PDS containing high-concentration glucose can induce significant apoptosis and inhibit the proliferation of RPMCs in vitro.</p>

Expression of SDF-1alpha and its receptor CXCR4 in acute leukemias and their relationship with extramedullary infiltration / 中国实验血液学杂志

The study was aimed to explore the expression of stromal cell derived factor-1alpha (SDF-1alpha) and its receptor CXCR4, and their relationship with the extramedullary infiltration in acute lymphoblastic, grannulocytic and monocytic leukemia. 66 cases of acute leukemia included 31 cases of acute lymphoblatic leukemia (ALL), 20 cases of acute grannulocytic leukemia (M(2)) and 15 cases of acute monocytic leukemia (M(4)+M(5)). There were 41 cases with extramedullary infiltration and 25 cases without-extramedullary infiltration. Enzyme-linked immunoabsorbent assay (ELISA) and flow cytometry were used to determine expression of SDF-1alpha and CXCR4 respectively on leukemia cells in peripheral blood and bone marrow of different groups. The results showed that average plasma level of SDF-1alpha in the ALL, M(4)+M(5), M(2) patients and the normal control were 1317.87 +/- 220.76, 1339.79 +/- 187.06, 1063.70 +/- 190.74, 1908.34 +/- 135.55 (pg/ml) respectively. The average levels in the ALL, M(4)+M(5) and M(2) patients groups were lower than those in normal control group. Both levels in ALL and M(4)+M(5) patient groups were higher than that in M(2) patient group. The average levels of SDF-1alpha in patient group with extramedullary infiltration and patient groups without-extramedullary infiltration were 1252.49 +/- 263.12, 1234.91 +/- 185.50 (pg/ml) respectively. The former seemed as if higher than the latter, but without statistical significance. The MFI of CXCR4 expression in ALL, M(4)+M(5), M(2) patient group were 78.47 +/- 33.96, 67.21 +/- 24.29, 41.66 +/- 17.18, respectively. CXCR4 expression in ALL and M(4)+M(5) patient groups were higher than that in M(2) patient group (P > 0.05). There was no significant difference between the ALL and M(4)+M(5) patient group (P > 0.05). The MFI of CXCR4 expression in patients with extramedullary infiltration and patients without extramedullary infiltration were 81.72 +/- 27.63, 36.94 +/- 11.86 respectively. The former was higher than the latter (P < 0.05). It is concluded that the higher expression of CXCR4 on acute lymphoblatic and monocytic leukemia cells may be one of the molecular mechanisms of extramedullary infiltration in both kinds of leukemia. The average plasma levels of SDF-1alpha decreased in leukemia patients and this decrease not related to the extramedullar infiltration, which may be due to the SDF-1alpha local expression in the organ infiltrated.

Clinical significance of antineutrophil cytoplasmic antibodies in patients with lupus nephritis / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze the clinical features of patients with lupus nephritis positive for antineutrophil cytoplasmic antibodies (ANCA) and explore the clinical implications of ANCA detection.</p><p><b>METHODS</b>Totally 261 patients with lupus nephritis were enrolled in this study, including 53 ANCA-positive and 208 ANCA-negative ones. The clinical data of the patients pertaining to the disease history, physical examination, laboratory examinations and pathological inspection were retrospectively analyzed.</p><p><b>RESULTS</b>Compared with patients negative for ANCA, the ANCA-positive patients had significantly higher incidence of serositis (75.5%), acute renal failure (64.2%), myocarditis (30.2%), neuropsychiatric involvement (26.4%) and lung hemorrhage (7.5%)(P<0.05). Significant differences were also found between the two groups in SLE disease active index (SLE-DAI), number of the diagnostic criteria, erythrocyte sedimentation rate (ESR), anemia, anti-Sm antibodies, and serum complement C(3). Most patients positive for ANCA (67.9%) had type IV lupus nephritis with more crescent formation, renal tubular atrophy, hyaline thrombi, and higher mortality rate as well than the negative patients.</p><p><b>CONCLUSION</b>ANCA detection may benefit the estimation of the disease severity and prognostic evaluation of lupus nephritis.</p>

CXCR4 expression in B-lineage acute lymphocyte leukemia and its significance / 中国实验血液学杂志

The study was aimed to explore the chemokine receptor CXCR4 expression on the B-lineage acute lymphocyte leukemia (B-ALL) cells of various differentiation stages and its relationship with myeloid antigen expression. Flow cytometry was used to detect the CXCR4 expression by means of double-fluorescence labeling with CD19/SCC gating. The results demonstrated that 92.9% B-ALL patients were positively expressed CXCR4. The CD10, CD34 antigens were differently expressed in differentiation stages of B-ALL. The immunotypes of (1) CD10(-)/CD34(+), (2) CD10(+)/CD34(+), (3) CD10(+)/CD34(-), (4) CD10(-)/CD34(-) presented at various differential stages from premature to mature. The positive rate of CXCR4 were (27.60 +/- 15.25)%, (30.95 +/- 15.50)%, (55.62 +/- 18.37)% and (77.25 +/- 10.86)% from (1) to (4) respectively. The median fluorescence intensity (MFI) of CXCR4 expression were 46.69 +/- 15.06, 47.43 +/- 12.39, 79.28 +/- 24.71 and 132.92 +/- 88.09. CXCR4 expressions were not significantly different between the premature stages of CD10(-)/CD34(+) and CD10(+)/CD34(+) subtypes, but both were lower than the CXCR4 expression in CD10(+)/CD34(-) and CD10(-)/CD34(-) subtypes. The highest incidence of CXCR4 expression was found in CD10(-)/CD34(-) B-ALL. The average level of CXCR4 expression on B-ALL cell with positive myeloid antigen CD13 or/and CD33 (my(+)B-ALL) was (12.56 +/- 3.88)% of positive rate and 39.82 +/- 11.58 of MFI, both of which were less than the positive rate (37.57 +/- 11.59)% and the MFI (50.72 +/- 13.34) on B-ALL cells with negative myeloid antigen expression (mye(-)B-ALL). In conclusion, the CXCR4 expression is associated with differentiation level of B-ALL cells and down-regulated when co-expressed with myeloid antigens.

TGF-?_1 induced the tolerant dendritic cells and the mechanism / 中华器官移植杂志

Objective To investigate the induction of tolerant dendritic cells(DOs)and its mechanism in mice.Methods The dendritic cell line derived from C57BL/6 mouse.DO2.4 cells were co-cultured with TGF-?_1(20 ng/ml)to induce the tolerant DCs(TGF?-DC)and stimulated wihh li- popolysaccharide(LPS)for 48 h to induce the mature DCs(LPS-DC).Special small interference RNA molecule(siRNA)of PIR-B was chemically synthesized and was transfected into TGF?-DC by lip2000 (si-DC).The expression of paired immunoglobin receptor A/B(PIR-A/B)in DC2.4 cells was mea- sured by semi-quantitative RT-PCR and flow cytometry(FCM).The allogeneic stimulating capacity of DCs was measured by mixed lymphocyte reaction(MLR)using ~3H-thymidine incorporation test with the Balb/c spleen cells as reaction cells.The concentration of IFN-?in supernatants of MLR from dis- tinct groups was tested by enzyme linked immunosorbent assay(ELISA).Results TGF-?_1 up-regula- ted the PIR-B mRNA expression and down-regulated the PIR-A mRNA expression(P