OBJECTIVE: To establish the in vivo traceable acute myeloid leukemia mice model with Luciferase-Expressing KG1a Cells. METHODS: KG1a cells with stable luciferase gene expression (called as KG1a-Luc cells) were constructed by lentivirus transfection, then sifted out by puromycin. Eighteen male NOD-SCID-IL2rg RESULTS: KG1a cells expressing luciferase stably were successfully obtained. The tumor luminescence wildly spread at day 17 captured by in vivo imaging. The KG1a-Luc tumor cells could be detected in the peripheral blood of the mice, with the average percentage of (16.27±6.66)%. The morphology and pathology result showed that KG1a-Luc cells infiltrate was detected in bone marrow, spleens and livers. The survival time of the KG1a-Luc mice was notably shorter as compared with those in the control group, the median survival time was 30.5 days (95%CI: 0.008-0.260). CONCLUSION The acute myeloid leukemia NOD-SCID-IL2rg
Animals ; Disease Models, Animal ; Interleukin Receptor Common gamma Subunit ; Leukemia, Myeloid, Acute ; Luciferases/genetics* ; Male ; Mice ; Mice, Inbred NOD ; Mice, SCID

Objective To compare the clinical efficacy and safety of tolvaptan tablets and furosemide injection in the treatment of heart failure with systolic dysfunction. Methods One hundred and four patients of heart failure with systolic dysfunction were randomly divided into control group (n = 49 cases) and treatment group (n = 55 cases) . Control group received furosemide 40 mg per time, qd, intravenous bolus. Treatment group received tolvaptan 15 mg per time, qd, orally. The clinical efficacy, heart rate, pulmonary artery pressure, pulmonary capillary pressure and cardiac output, and adverse drug reactions were compared between two groups. Results After treatment, the total effective rates of treatment and control groups were 89. 09％ (49 cases/55 cases) and71. 43％ (35 cases/49 cases) with significant difference (P < 0. 05) .After treatment, the main indexes of treatment and control groups were compared: heart rates were (80. 15 ± 10. 04) and (84. 71 ± 9. 66) beat·min-1, pulmonary artery pressure were (21. 85 ± 4. 49) and (28. 47 ± 4. 46) mm Hg, pulmonary capillary pressure were (11. 24 ± 1. 61) and (15. 18 ± 2. 76) mm Hg, cardiac output were (1. 94 ± 0. 30) and (2. 16 ± 0. 25) L · min-1· m-2, the differences were statistically significant (all P < 0. 05) . The adverse drug reactions of treatment group were increased blood sodium, urinary frequency and dry mouth, which in control group were fatigue, thirst and muscle soreness. The total incidences of adverse drug reactions in the treatment and control groups were 12. 73％ and 12. 24％ without significant difference (P> 0. 05) . Conclusion Tolvaptan tablets have a definitive clinical efficacy in the treatment of heart failure with systolic dysfunction, which can effectively improve the heart rate, pulmonary artery pressure, pulmonary capillary pressure and cardiac output, without increasing the incidence of adverse drug reactions.

OBJECTIVETo investigate the pathogenesis of ischemic-type biliary lesions (ITBLs) in post-liver transplant patients and the possible therapeutic mechanisms of sirolimus.

METHODSThe clinic data of 32 post-liver transplant patients with ITBLs from May 2004 to December 2010 was analyzed. There were including 25 male and 7 female patients with a median age of 46 years (ranging from 19 to 61 years). Patients were divided into those who received sirolimus (sirolimus group) and those who did not (control group). The expression of IL-2, FoxP3, and IL-10 in the portal area, liver function indexes, and bile duct injury score were assessed pre-ITBL, when ITBLs were identified, and after 6 months of sirolimus treatment.

RESULTSCompared with pre-ITBL optical density (OD) values, there was a significantly increase in IL-2 OD(0.138 ± 0.050 in control group and 0.141 ± 0.052 in sirolimus group), but not FoxP3 and IL-10 OD in both groups at the time ITBLs were diagnosed. After 6 months of treatment, the IL-2, FoxP3, and IL-10 OD values in the control group were not different from those when ITBLs were diagnosed. There was a significant reduction in post-therapy IL-2 OD(0.107 ± 0.043, t = 2.087, P = 0.044), and a significant elevation in FoxP3(0.213 ± 0.039) and IL-10 OD(0.187 ± 0.048) in sirolimus group as compared with those when ITBLs were diagnosed(t = -3.822 and -4.350, both P < 0.01). There was a significant increase in serum levels of ALT, AST, total bilirubin, γ-glutamyl transpeptidase and ALP at the time ITBLs were diagnosed compared with pre-ITBL levels in both groups. After 6 months of treatment, the above indexes had not changed in the control group, but significantly improved in the sirolimus group, and the bile duct injury score in the sirolimus group had significantly decreased(4.4 ± 2.4, Z = -2.568, P = 0.010). The 1-year and 3-year graft survival rates in the control group were 6/13 and 5/13, respectively, and 17/19 and 13/19, respectively, in the sirolimus group (χ(2) = 7.166, P = 0.007; χ(2) = 5.398, P = 0.020, respectively).

CONCLUSIONSSirolimus can downregulate IL-2 expression and upregulate FoxP3 and IL-10 expression, thereby stimulating FoxP3+ Treg cells, suppressing immunopathological damage, and promoting epithelial repair in bile ducts.

Adult ; Bile Duct Diseases ; drug therapy ; Female ; Forkhead Transcription Factors ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Ischemia ; diet therapy ; Liver Transplantation ; Male ; Middle Aged ; Postoperative Complications ; drug therapy ; Sirolimus ; therapeutic use ; Young Adult

OBJECTIVETo explore the effectiveness of the traditional Chinese herbal medicine Lingzhi (Ganoderma lucidum) preparation in treating simian acquired immune deficiency syndrome (SAIDS).

METHODSFive female adult Chinese rhesus monkeys were inoculated rectally with SIVmac239, and were all diagnosed as SAIDS by laboratory and clinical examinations 17 months later. Of these 5 monkeys, 3 (#393, #374, and #381; treatment group) were orally administered with Ganoderma lucidum (2 spores powder capsules plus 2 spores oil capsules on a daily basis), and the remaining other two monkeys (#348 and #361) served as control and did not receive treatment.

RESULTSAnimal #393 (treatment group), #361 (control group) and #348 (control group) died of SAIDS (opportunity infection) 3.5 months, 6 months, and 11 months later, respectively. Two animals (#374 and #381) survived. The necropsy revealed depletion and/or exhaustion of their lymphoid tissue. In the monkey #374, the peripheral CD4(+) T lymphocyte increased by 30% in the 6(th) month compared with the baseline level and then fluctuated. The plasma viral load gradually fell and reached about 1 log(10) in the treatment group, but remained stable in the control group. As shown by pathological examinations, the lymph node and spleen of monkeys #374 (treatment group) and #381 (treatment group) showed rehabilitation and reconstruction in the lymphatic tissue, thymus, nerve tissue of gyrus hippocampi, pituitary gland, pineal body, thyroid gland, adrenal gland, and ovary. In the control group, however, animals experienced depletion of lymph nodes, atrophy of spleen, disappearance of thymus, and other disorders in endocrine organs.

CONCLUSIONGanoderma lucidum preparation may have certain protective effect on the immune system, nervous system, and endocrine system of monkeys with SAIDS.

Animals ; Disease Models, Animal ; Female ; Macaca mulatta ; Materia Medica ; therapeutic use ; Reishi ; Simian Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Treatment Outcome

Adult ; Carcinoma, Hepatocellular ; epidemiology ; pathology ; virology ; DNA, Viral ; blood ; genetics ; Female ; Genotype ; Hepatitis B ; epidemiology ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; epidemiology ; pathology ; virology ; Male ; Middle Aged ; Neoplasm Staging ; Polymerase Chain Reaction ; methods

OBJECTIVETo observe the effects of different cell lysis buffers on protein quantification with Bradford method and bicinchoninic acid (BCA) method.

METHODSBradford method and BCA method were used to determine the concentration of bovine serum albumin (BSA) in different solutions (distilled water, cell lysis buffer used in two-dimensional differential in-gel electrophoresis and three kinds of cell lysis buffers used in conventional two dimensional gel electrophoresis), as well as the protein concentrations of cell lysates using these different lysis buffers. Bradford method was also applied to determine the protein concentrations of samples with repeated freeze thaw cycle, in different colorimetric cylinders, or using different standard curves from different periods.

RESULTThe protein measurements increased for 1.2 to 2 fold when different cell lysis buffers were used in Bradford method, but the measurements increased with the increased concentration of BSA (r=0.989 approximately 0.996, P<0.05). For BCA, measurement reading increased about thousands times higher, even overflowed the limits of machine. Protein measurements didn't change significantly, only showed a declined trend after repeated freeze thaw cycle, while no significant changes were found using different colorimetric cylinders or standard curves from different periods.

CONCLUSIONBradford method may be the choice of the protein quantification in proteomics. However, optimization is required for specific experimental conditions.

Buffers ; Cells ; Chemistry Techniques, Analytical ; methods ; Proteins ; analysis ; Serum Albumin, Bovine ; analysis ; Spectrophotometry, Ultraviolet

OBJECTIVETo investigate the biological and clinical features of Chinese rhesus monkeys after intravenous (IV) and intrarectal (IR) challenge with SIVmac239 in rhesus monkeys of Chinese origin, and compare the differences between the routes of infection.

METHODSRhesus monkeys of Chinese origin were inoculated with SIVmac239 either by IV (n = 19) or IR (n = 6) routes. Simian immunodeficiency virus (SIV)-specific antibody titer, CD4 + T cell counting, plasma SIV load, lymph node pathology, and clinical manifestations were compared between these two groups 232 or 168 days after challenging.

RESULTSAll SIVmac239-inoculated animals became seropositive for SIV-specific antibodies. SIV-specific IgM was detected in IV groups as from day 10 but was not detected in IR for all the time points. Although SIV-specific IgG was detected as from day 30 in both groups, the IgG titers were ten-fold higher in IV group than in IR group after day 168. CD4 + T-cell counting decreased progressively in IV group but remained stable in IR group over time. Plasma SIV RNA loads peaked in all animals between day 10 and day 14 (10(7) copies/ml), then declined to "setpoint" (10(3) - 10(6) copies/ml) about 2 months later. Most inoculated animals manifested lymphadenopathy. Two animals in IV group and one in IR group died of simian AIDS between day 150 and day 210, as evidenced by the autopsies showing the depletion of lymph tissues, Pneumocystis carinii pneumonia and other opportunity infections. Conclusion IV or IR inoculation of SIVmac239 in Chinese rhesus monkeys will result in chronic SIV infection with a similar clinical feature of natural HIV infection, which provides an excellent experimental animal model for AIDS.

Animals ; CD4-Positive T-Lymphocytes ; metabolism ; China ; Disease Models, Animal ; Female ; Macaca mulatta ; virology ; Male ; Rectum ; virology ; Simian Acquired Immunodeficiency Syndrome ; immunology ; virology ; Simian Immunodeficiency Virus ; immunology ; pathogenicity ; Veins ; virology

OBJECTIVENo optimal prophylactic protocol of hepatitis B immunoglobulin (HBIG) combined with nucleos(t)ide analogue for HBV recurrence has been established yet. By investigating the alterations of HBV markers in HBV related liver disease patients, recipients of a liver transplant, under lamivudine or/and HBIG prophylaxis, we aim to explore the possible HBV recurrence mechanism involved and to find a new option in the prophylaxis of HBV recurrence and to tailor individualized therapy.

METHODSSerial liver biopsy specimens and sera were obtained intraoperationally and at definite time points during follow-up. ELISA and chemiluminescent microparticle immunoassay, HBV DNA fluorescent quantification, immunohistochemistry staining and HBV DNA in situ hybridization were performed. Alterations of HBV markers in specimens of 96 liver transplant recipients were investigated retrospectively.

RESULTSAll 17 cases had HBV recurrence (median 37 months) which occurred in the follow-up period after liver transplantation. The overall actual HBV recurrence rate at 2 years was 22% with a significant difference between that of the active and inactive groups (P<0.05); 82.4% HBV recurrence took place within the first 3 years after the operation, and the recurrence ratio of first 3 years to 3 years later after transplantation was 4.7 (P<0.01). The HBV DNA positive patients accounted for 78.6% of the total number of recurrences within the first 3 years. HBcAb and HBeAb positive rates went down with time, but their positivity remained.

CONCLUSIONHBV recurrence happens after liver transplantation. In inactive HBV replicative patients with strictly combined prophylaxis and availability of other medications and using 3 years after liver transplantation as a point of time, we think that tapering down the dosage of HBIG and tailoring individualized treatment methods based on virological and immunological situations of each recipient are worth trying.

Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Child ; Female ; Hepatitis B ; drug therapy ; Humans ; Immunoglobulins ; therapeutic use ; Lamivudine ; therapeutic use ; Liver Transplantation ; Male ; Middle Aged ; Postoperative Period ; Prognosis ; Retrospective Studies ; Secondary Prevention ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the expression of angiotensin converting enzyme 2 (ACE2) and the changes treated with angiotensin converting enzyme inhibitor (ACEI), and its signal transduction pathway.

METHODSAtrial tissues were obtained from 47 patients with RHD undergoing cardiac surgery. The mRNA of ACE2 and ACE were semi-qualified by RT-PCR and normalized to the gene beta-actin. Western blot analysis was employed to examine the expressions of ACE2, ACE, ERK1/2 and phosphorylated ERK (pERK1/2). The atrial tissue angiotensin II (Ang II) content was determined by radioimmunoassay detection.

RESULTSThe expression of ACE2 was significantly decreased (P < 0.05), the expression of ACE and pERK1/2 were significantly increased (P < 0.05), and the level of atrial tissue Ang II was significantly increased in patients with chronic atrial fibrillation group (CAF) compared with sinus rhythm group (SR) (P < 0.05). Compared with CAF patients treated without ACEI, the expression of ACE2 significantly increased (P < 0.01), and the relative activity of ERK1/2 significantly decreased (P < 0.05), whereas the expression of ACE and the level of atrial tissue Ang II remained unchanged in CAF patients treated with ACEI.

CONCLUSIONSThe study suggested that the dysequilibrium of ACE/ACE2 might play an important role in the process of atrial fibrillation, which may be related to the activation of ERK1/2 pathway. The clinical effect of long-term treatment of ACEI maybe associated with elevated ACE2 expression but not ACE expression.

Adult ; Aged ; Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Atrial Fibrillation ; drug therapy ; metabolism ; Female ; Heart Atria ; metabolism ; Humans ; Male ; Middle Aged ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Peptidyl-Dipeptidase A ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction

OBJECTIVETo study the relationship between simian acquired immunodeficiency syndromn (SAIDS) and autoimmunity in simian immunodeficiency virus (SIV)-infected monkeys.

METHODSIndirect immunofluorescence assays were performed to detect plasma or serum autoantibodies in SIV-infected monkeys. The heart, liver, spleen, lung, kidney, and lymph node of BALB/c mice, a strain of endothelial cell ECV304, and granulocytes were used as target antigens. These results were compared with HE stained slides of SIV-infected monkeys.

RESULTSThe levels of various autoantibodies, including anti-lymphocyte autoantibodies, anti-endothelial cell autoantibodies, and anti-granulocyte antibodies, increased after SIV infection in monkeys. Moreover, pathological examinations showed injuries in the lymphoid tissue and vascular pathological changes in cerebral cortex, submucosa of gastrointestinal tract, interstitial capillaries of myocardium, nephron of the kidney, and sinusoid cell of liver.

CONCLUSIONThe increased autoantibodies and the pathological changes of tissues and organs confirm the existence of autoimmunity in SIV-infected monkeys.

Animals ; Autoantibodies ; blood ; Autoimmunity ; Endothelial Cells ; immunology ; Granulocytes ; immunology ; Lymphocytes ; immunology ; Mice ; Mice, Inbred BALB C ; Simian Acquired Immunodeficiency Syndrome ; immunology ; pathology ; Simian Immunodeficiency Virus