Clinical characteristics of rheumatic disease-associated hypophysitis: A case series and review of literature.

Rheumatic diseases have been reported to sometimes involve the pituitary gland. This study aims to characterize the clinical features and outcomes of patients with rheumatic disease-associated hypophysitis. We used the electronic medical record system in our hospital to identify nine patients with pituitary involvement in rheumatoid disease. We summarized the clinical characteristics, radiographic findings, treatments, and clinical outcomes of the 9 patients. We also performed a systematic literature review of systemic lupus erythematosus (SLE) cases with pituitary involvement published in PubMed and Wanfang databases from 1995 to 2021, and eight patients with complete information were selected. In the nine-patient cohort, the median age was 54 years, and the spectrum of rheumatic diseases included immunoglobulin G4-related disease (IgG4RD) (4/9), SLE (2/9), vasculitis (2/9), and Sjögren syndrome (SS) (1/9). All patients had pituitary abnormalities on radiological assessment, 6 developed diabetes insipidus (DI), and 8 presented with anterior pituitary hormone deficiencies in the disease duration. All the patients had multisystem involvement. As compared to hypophysitis with IgG4RD (IgG4-H), the age at onset of hypophysitis with SLE (SLE-H) patients was younger [(30.4 ± 16.4) years vs. (56.0 ± 0.8) years] and the disease duration was shorter [(14.0 ± 17.5) months vs. (71.0 ± 60.9) months] (P < .05). All patients were managed with glucocorticoids (GC) in combination with another immunosuppressant, and the majority of patients improved within 4 months. Six patients achieved disease remission while four required at least one hormone replacement therapy. Hypophysitis is a rare complication secondary to a variety of various rheumatic diseases that can occur at any stage. GC combined with additional immunosuppressants could improve patients' symptoms; however some patients also required long-term hormone replacement therapy in pituitary disorders.

Antioxidant Peptides From Protein Hydrolysate of Marine Red Algae Eucheuma cottonii: Preparation, Identification, and Cytoprotective Mechanisms on H2O2 Oxidative Damaged HUVECs.

To screen, prepare, identify, and evaluate the activities of natural antioxidants for treating chronic diseases caused by oxidative stress. Two algal proteins, namely ZD10 and ZD60, precipitated with 10 and 60% (NH4)2SO4 were extracted from red algae Eucheuma cottonii (E. cottonii) and hydrolyzed using five proteolytic enzymes. The results showed that ZD60 played the most significant role in the enhancement of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH⋅) scavenging activity (25.91 ± 0.24%) among all protein hydrolysates. Subsequently, six antioxidant peptides (EP1-EP6) were isolated from the papain hydrolysate of ZD60 by ultrafiltration and chromatography methods. Their amino acid sequences were identified as Thr-Ala (EP1), Met-Asn (EP2), Tyr-Ser-Lys-Thr (EP3), Tyr-Ala-Val-Thr (EP4), Tyr-Leu-Leu (EP5), and Phe-Tyr-Lys-Ala (EP6) with molecular weights of 190.21, 263.33, 497.55, 452.51, 407.51, and 527.62 Da, respectively. Of which, EP3, EP4, EP5, and EP6 showed strong scavenging activities on DPPH⋅, hydroxyl radical (HO⋅), and superoxide anion radical (O- 2⋅). Moreover, EP4 and EP5 could significantly protect human umbilical vein endothelial cells (HUVECs) from H2O2-induced oxidative damage by increasing the levels of antioxidant enzyme systems including superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) to reduce the levels of reactive oxygen species (ROS) (60.51 and 51.74% of model group) and malondialdehyde (MDA) (75.36 and 64.45% of model group). In addition, EP4 and EP5 could effectively inhibit H2O2-induced apoptosis by preventing HUVECs from early apoptosis to late apoptosis. These results indicated that the antioxidant peptides derived from E. cottonii, especially EP4 and EP5, could serve as the natural antioxidants applied in pharmaceutical products to treat chronic cardiovascular diseases caused by oxidative damage, such as coronary heart disease, atherosclerosis, etc.

The NF-κB signalling pathway and TM7SF3 contribute to liver fibrosis caused by secreted phospholipase A2 of Clonorchis sinensis.

BACKGROUND: The NF-κB signalling pathway has been reported to be related to liver fibrosis, and we investigated whether the NF-κB signalling pathway is involved in liver fibrosis caused by secreted phospholipase A2 of Clonorchis sinensis (CssPLA2). Furthermore, expression of the receptor of CssPLA2 on the cell surface of hepatic stellate cells (HSCs) may greatly contribute to liver fibrosis. METHODS: CssPLA2 was administered to BALB/c mice by abdominal injection. The levels of markers of NF-κB signalling pathway activation in mouse liver tissue were measured by quantitative RT-PCR, ELISA and western blot. Additionally, HSCs were incubated with CssPLA2, and an NF-κB signalling inhibitor (BAY 11-7082) was applied to test whether the NF-κB signalling pathway plays a role in the effect of CssPLA2. Then, the interaction between CssPLA2 and its receptor transmembrane 7 superfamily member 3 (TM7SF3) was confirmed by co-immunoprecipitation (co-IP) and GST pull-down. To determine how TM7SF3 influences the ability of CssPLA2 to cause liver fibrosis, a TM7SF3 antibody was used to block TM7SF3. RESULTS: The levels of the NF-ΚB signalling pathway activation markers TNF-α, IL-1ß and phospho-p65 were increased by CssPLA2 in the context of liver fibrosis. In addition, the interaction between TM7SF3 and CssPLA2 was confirmed by co-IP and GST pull-down. When TM7SF3 was blocked by an antibody targeting 1-295 amino acids of TM7SF3, activation of HSCs caused by CssPLA2 was alleviated. CONCLUSIONS: The NF-ΚB signalling pathway is involved in the activation of HSCs by CssPLA2. TM7SF3, the receptor of CssPLA2, plays important roles in liver fibrosis caused by CssPLA2.

High Fischer ratio oligopeptides determination from Antartic krill: Preparation, peptides profiles, and in vitro antioxidant activity.

In this work, alcalase and flavorzyme were chosen as the hydrolase for preparing high Fischer ratio oligopeptides from Antarctic krill (Euphausia superba) (HFP) using sequential enzyme hydrolysis process, and their hydrolysis conditions were optimized using single factor experiment. According to the Fischer ratio, granular activated carbon of XHJ-200 (200 mesh) showed the best way to remove aromatic amino acids and its optimal parameters were pH value of 6.0, adsorption time of 2.5 hr, temperature of 25°C, and solid-liquid ratio of 1:20. The Fischer ratio and average molecular weight of HFP were 21.12 (>20) and 779.9 Da, respectively. In addition, the peptide profile of HFP was established using RP-HPLC and 23 oligopeptides isolated from HFP including 6 dipeptides, 9 tripeptides, 3 tetrapeptides, and 5 pentapeptides were identified using protein amino acid sequence analyzer and mass spectrum. Furthermore, HFP exhibited high radical scavenging activity, reducing power, and lipid peroxidation inhibition capability. PRACTICAL APPLICATIONS: High Fischer ratio oligopeptides (HFO) is a kind of small peptide mixture with the mole ratio of branched-chain amino acids (BCAA) to aromatic amino acids (AAA) higher than 20, which has drawn a great attention due to its great potential in clinical nutrition approaches for the treatment of liver disease when amino acid composition is out of proportion characterized by low levels of BCAA and high levels of AAA in the systemic blood. Antarctic krill is regarded as the largest animal protein resource for various food and pharmaceutical products. However, there is no report on the preparation of HFO of Antarctic krill (HFP). Therefore, the aim of the present study was to investigate the preparation process, peptide profiles and in vitro antioxidant activity of HFP. This study will potentially enhance the value-added utilization of Antarctic krill by making it an important raw material in the health-promoting functional products.

[Effect of ultraviolet irradiation on the proliferation of acute promyelocytic leukemia cells under hypoxic conditions and related mechanisms].

OBJECTIVE: To study the effect of 280 nm-LED ultraviolet irradiation on the proliferation of acute promyelocytic leukemia (APL) HL-60 cells under hypoxic conditions and related mechanism. METHODS: HL-60 cells in the logarithmic growth phase were selected and divided into control, hypoxia, ultraviolet and hypoxia+ultraviolet groups. The cells in the hypoxia group were treated with cobalt chloride (with a final concentration of 150 µmol/L), those in the ultraviolet group were irradiated by 280 nm-LED ultraviolet with an energy intensity of 30 J/m2, and those in the hypoxia+ultraviolet group were treated with cobalt chloride and then irradiated by 280 nm-LED ultraviolet. After 48 hours of treatment, the cells were placed under an invert microscope to observe cell morphology. CCK-8 assay was used to measure the inhibition rate of cell proliferation. Annexin V-FITC/PI double staining flow cytometry was used to evaluate cell apoptosis. Quantitative real-time PCR was used to measure the mRNA expression of Bcl-2. Each experiment above was repeated three times independently. RESULTS: Compared with the control group, the experimental groups showed shrinkage, decreased brightness, and disordered arrangement of cells, and the number of cells decreased over the time of culture. There were significant differences in the inhibition rate of cell proliferation and cell apoptosis rate among the groups (P<0.01), and the hypoxia+ultraviolet group showed the strongest inhibition of cell proliferation and induction of cell apoptosis, followed by the ultraviolet group and the hypoxia group. Compared with the control group, the other three groups had a gradual reduction in the mRNA expression of Bcl-2, and the hypoxia+ultraviolet group had a significantly greater reduction than the hypoxia and ultraviolet groups (P<0.01). CONCLUSIONS: Both hypoxia and ultraviolet irradiation can inhibit the proliferation of HL-60 cells and induce cell apoptosis, and ultraviolet irradiation has a better effect on proliferation inhibition and cell apoptosis under hypoxic conditions than under normoxic conditions, possibly by downregulating the mRNA expression of Bcl-2.

Purification and Identification of Antioxidant Peptides from Protein Hydrolysate of Scalloped Hammerhead (Sphyrna lewini) Cartilage.

The aim of this study was to purify and identify peptides with antioxidant properties from protein hydrolysate of scalloped hammerhead (Sphyrna lewini) cartilage. Cartilaginous proteins of the scalloped hammerhead were extracted by guanidine hydrochloride, and three antioxidant peptides, named enzymolysis peptide of scalloped hammerhead cartilage A (SCPE-A), SCPE-B and SCPE-C, were subsequently isolated from the hydrolysate of the cartilaginous proteins using ultrafiltration and chromatography. The amino acid sequences of SCPE-A, SCPE-B and SCPE-C were identified as Gly-Pro-Glu (GPE), Gly-Ala-Arg-Gly-Pro-Gln (GARGPQ), and Gly-Phe-Thr-Gly-Pro-Pro-Gly-Phe-Asn-Gly (GFTGPPGFNG), with molecular weights of 301.30 Da, 584.64 Da and 950.03 Da, respectively. As per in vitro activity testing, SCPE-A, SCPE-B and SCPE-C exhibited strong scavenging activities on 2,2-diphenyl-1-picrylhydrazyl radicals (DPPH•) (half elimination ratio (EC50) 2.43, 2.66 and 1.99 mg/mL), hydroxyl radicals (HO•) (EC50 0.28, 0.21 and 0.15 mg/mL), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radicals (ABTS⁺•) (EC50 0.24, 0.18 and 0.29 mg/mL), and superoxide anion radicals ( O 2 - •) (EC50 0.10, 0.14 and 0.11 mg/mL). In addition, SCPE-A showed inhibition activity similar to butylated hydroxytoluene (BHT) in lipid peroxidation in a linoleic acid model system. The amino acid residues of Gly, Pro and Phe could positively influence the antioxidant activities of GPE, GARGPQ and GFTGPPGFNG. These results suggested that GPE, GARGPQ and GFTGPPGFNG might serve as potential antioxidants and be used as food additives and functional foods.

Clinical presentation and outcome of pediatric patients with hemophagocytic lymphohistiocytosis in China: A retrospective multicenter study.

BACKGROUND: Hemophagocytic lymphohistiocytosis (HLH) is a heterogeneous disease with major diagnostic and therapeutic difficulties. A large-scale multicenter study of pediatric HLH is still lacking in China. PROCEDURE: The Histiocytosis Study Group of the Chinese Pediatric Society conducted this retrospective study in 2014. A total of 323 patients diagnosed with HLH between 2011 and 2013 from 12 hospitals were registered. RESULTS: The median age at diagnosis was 2.2 years (range, 0-14.6 years), with a peak age of HLH onset at 0 to 3 years (63%). Mutations in HLH-related genes were found in 27.9% (24/86) patients who underwent genetic testing. PRF1, UNC13D, STXBP2 and LYST were the predominant genes involved. Sixteen patients (66.7%) presented with only monoallelic mutations in one gene. Epstein-Barr virus (EBV) infection was the major condition related to HLH, which was documented in 74.4% (201/270) of the patients who underwent EBV detection. Of 252 evaluable patients, 64.7% (163) achieved non-active disease at the eighth week and patients treated with a protocol containing etoposide presented higher remission rates (75.6% vs. 46.8%, P < 0.001). In multivariate analysis, a younger age at diagnosis (<12 months), platelet count less than 80×109 /L, central nervous system involvement, and initial treatment using a protocol without etoposide (not HLH-94/04) were independent prognostic factors indicating resistant disease. DISCUSSION: This study first multicenter assessment of HLH in China shows some different features in Chinese children with HLH compared with those in western countries, including older age, vulnerability to EBV infection, and a high proportion of patients with single monoallelic genetic mutations.

[Expression of PTEN in Myocardial Tissue in Coronary Heart Disease].

OBJECTIVE: To observe the expression of phosphatase and tensin homology deleted on chromosome ten (PTEN) in myocardial tissue in patients with coronary heart disease, and explore the relevance between the expression of PTEN and the occurrence and development of coronary heart disease. METHODS: A total of 16 death cases with pathological diagnosis of coronary heart disease were collected as experimental group, and 19 cases without myocardial lesions were selected as control group. The expression of PTEN protein and its mRNA were detected by immunohistochemistry and real-time fluorescence quantitative PCR respectively. The correlation between the expression of PTEN and the pathogenesis of coronary heart disease was analyzed. RESULTS: The expression of PTEN protein in myocardium in cases with coronary heart disease was significantly lower compared with the control group (P < 0.05). There was no statistical difference of the expression of PTEN mRNA between experimental and control group (P > 0.05). CONCLUSION: PTEN may be involved in the occurrence and development of coronary heart disease.

[Relationship between clinical features and cognitive function in patients with childhood and adolescence-onset schizophrenia].

OBJECTIVE: To explore the factors influencing cognitive functions in patients with childhood and adolescence-onset schizophrenia. METHODS: The clinical data of 78 patients with childhood and adolescence-onset schizophrenia who met with the criteria of ICD-10 for schizophrenia were retrospectively reviewed. The cognitive functions were evaluated by the Chinese Wechsler Intelligence Scale for Children (C-WISC), the Wisconsin Card Sorting Test (WCST), digit span backward and P300. The clinical symptoms were evaluated by the Positive and Negative Syndrome Scale (PANSS). RESULTS: The patients with a lower education level or earlier onset of age had a longer P3 latency at the P300Fz area. The patients with a higher parental education level had higher scores of full intelligence quotient (FIQ), verbal intelligence quotient (VIQ), performance intelligence quotient (PIQ), conceptual level and completed categories of WCST and backward numeric order reciting. The patients with higher PANSS negative subscale scores had lower scores of FIQ, VIQ, PIQ, completed categories and conceptual level of WCST and backward numeric order reciting. The patients with a longer stabilization time had higher backward numeric order reciting scores. CONCLUSIONS: The severity of negative symptoms of the patients and the educational level of their parents are major factors influencing cognitive functions in patients with childhood and adolescence-onset schizophrenia.

[Construction of Plasmodium falciparum signal peptide peptidase-GFP mutant and its expression analysis in the malaria parasite].

OBJECTIVE: To construct recombinant plasmid pSPPcGT which contains signal peptide peptidase gene of Plasmodium falciparum (PJSPP) and GFP, and transfect into P. falciparum (3D7 strain) to obtain mutant parasites which can express PJSPP-GFP. METHODS: Plasmodium falciparum(3D7 strain) genomic DNA was extracted from cultured malaria parasites. The C-terminal region of PJSPP, an 883 bp gene fragment was amplified by PCR, and then cloned into pPM2GT vector to get recombinant vector pSPPcGT. The recombinant vectors were identified by PCR, double restriction enzyme digestion and DNA sequencing. pSPPcGT vector was transfected into malaria parasites. The positive clones were selected by adding inhibitor of Plasmodium falciparum dihydrofolate reductase WR99210 to the culture medium. The pSPP-GFP-transfected parasites were fixed with methanol, stained with DAPI, and observed under immunofluorescence microscope. The PJSPP-GFP expression in P. falciparum was identified by SDS-PAGE and Western blotting. RESULTS: The C-terminal region of PJSPP was amplified from P.falciparum (3D7 strain) genomic DNA by PCR with the length of 883 bp. The constructed recombinant vectors were identified by PCR screening, double restriction enzyme digestion and DNA sequencing. The pSPPcGT vector was transfected into P. falciparum and the positive clones were selected by WR99210. GFP fluorescence was observed in transfected parasites by immunofluorescence microscopy, and mainly located in the cytoplasm. The PJSPP-GFP expression in malaria parasites was confirmed by Western blotting with a relative molecular mass of Mr 64,000. CONCLUSION: Recombinant vector PJSPP-GFP is constructed and transfected into P. falciparum to obtain P. falciparum mutant clone which can express PfSPP-GFP.

[Effect of parent training in combination with methylphenidate treatment on family relationships for children with attention deficit/hyperactivity disorder].

OBJECTIVE: To investigate the effect of parent training combined with methylphenidate treatment on family relationships in children with attention deficit/hyperactivity disorder (ADHD). METHODS: Fifty-nine parents of children with ADHD under methylphenidate treatment participated in a modified 5-week training program. The intervention effect was evaluated using the Conners Parent Symptom Questionnaire, ADHD Rating Scale-IV Home Version (ADHD-RS-IV Home Version), Caregiver Strain Questionnaire, Parent-Child Relationship Self-rating Scale and Piers-Harris Children's Self-Concept Scale. Parents also completed the training satisfaction survey before and after the intervention. RESULTS: After the 5-week parent training, compared with the baseline values, total scores of Conners Parent Symptom Questionnaire and scores of conduct problems and anxiety significantly decreased, and scores of attention deficit, hyperactivity, impulsivity and oppositional defiant behaviors of ADHD-RS-IV Home Version, and Caregiver Strain Questionnaire total scores were all significantly decreased (P<0.05), while total scores of the Parent-Child Relationship Self-Rating Scale and Piers-Harris Children's Self-Concept Scale were significantly increased (P<0.05). CONCLUSIONS: Modified 5-week parent training program may improve parent-child relationship and reduce parenting stress in ADHD families.

[Study on attention-deficit hyperactivity disorder in children based on resting-state functional magnetic resonance imaging].

OBJECTIVE: To compare resting-state functional magnetic resonance imaging (fMRI) findings of children with attention-deficit hyperactivity disorder (ADHD) and normal children, and to investigate the possible mechanism of brain dysfunction in children with ADHD. METHODS: Resting-state fMRI was performed on 18 children who met the DSM-IV diagnostic criteria for ADHD (ADHD group) and 18 normal children (control group) matched for age, sex, IQ, degree of education and handedness. The two groups were compared in terms of amplitude of low frequency fluctuation (ALFF) and regional homogeneity (ReHo). RESULTS: Compared with the control group, the ADHD group had decreased ALFF in the bilateral posterior lobes of the cerebellum and the left side of the pons, increased ALFF in the right precentral gyrus, decreased ReHo in the left medial frontal gyrus, right superior frontal gyrus, and left precuneus, and increased ReHo in the left anterior lobe of the cerebellum, left caudate nucleus, right parahippocampal gyrus, left precentral gyrus, and right middle frontal gyrus. CONCLUSIONS: In resting state, children with ADHD have decreased brain activity in some regions, including the cerebellum and frontal cortex, compared with normal children, which supports the hypothesis of dysfunctional fronto-cerebellar circuits in ADHD.

[Influential factors for methylphenidate treatment compliance in children with attention deficit hyperactivity disorder].

OBJECTIVE: To investigate influential factors for the tendency to medicate and medication compliance in children with attention deficit hyperactivity disorder (ADHD). METHODS: A total of 188 children aged from 5 to 16 years, who were initially diagnosed with ADHD according to DSM-IV criteria, were included in the study. They underwent symptom assessment and cognitive function test. The compliance of methylphenidate treatment was evaluated. RESULTS: Patients with better emotional state, and fewer oppositional and hyperactive behaviors and those who had a family history of psychiatric diseases and who obtained lower scores in the number cancellation test (NCT), were more prone to medication and/or exhibited better medication compliance. Logistic regression analysis showed that fewer oppositional and hyperactive behaviors and lower NCT scores were the predictive factors for a higher tendency to medicate, and a better emotional state was the predictive factor for better medication compliance. Patients of predominantly inattentive type were more prone to medication and showed better medication compliance, as compared with those of combined type. Gender, age and symptom severity were not associated with the tendency to medicate and/or medication compliance. CONCLUSIONS: There is a need to enhance medication compliance in children with ADHD who have hyperactive, impulsive and oppositional behaviors, and to improve their long-term social functions.

[Influence of crocin on proliferation in vitro and function of dendritic cells derived from bone marrow of children with acute leukemia].

This study was purposed to investigate the effect of crocin on the proliferation in vitro and immune function of dendritic cells (DC) derived from the bone marrow of children with acute leukemia. The mononuclear cells were isolated from bone marrow of leukemia children by Ficoll-Hypaque. The experiment was divided into six groups: blank control group (A), crocin 1.25 mg/ml group (B), cytokines (rhGM-CSF 75 ng/ml+rhIL-4 75 ng/ml+rhTNF-α 50 ng/ml) group (C), cytokines+crocin 0.3125, 1.25 or 5.0 mg/ml groups (D, E, F). The numbers of DC were counted and the phenotypes of DC were determined by flow cytometry on the ninth day of culture. The DC of different groups were mixed with T cells just separated from peripheral blood of another children with acute lymphoblastic leukemia, and cultured with rhIL-2 200 U/ml for 5 d. The function of DC was detected by mixed lymphocyte reaction (MLR). The results indicated that the test groups and control group all obtained a certain amount of typical DC, but the DC numbers in test groups were all higher than those in control group (P < 0.01). Cultured for 9 days, the rates of CD1a(+), CD83(+), and HLA-DR(+) in group C, D, E, F were higher than group A (P < 0.01). There was no statistically significant difference between A and B groups (P > 0.05). MLR showed that with the increasing of DC, the stimulation index of T cells in group A and B was not rising (P > 0.05); the stimulated index of T cells in group C and E was significantly rising, there was statistically significant difference between them (P < 0.01). When the number of stimulated cells was the same, the stimulation index of T cell in group E was the highest (P < 0.01). It is concluded that the capability of DC proliferation promoted by crocin alone is lower than that of its combination with rhGM-CSF, rhIL-4 and rhTNF-α, but the crocin can synergically promote the maturity of DC cooperating with rhGM-CSF, rhIL-4 and rhTNF-α. The DC induced by crocin can particularly enhance the proliferation of T cells.

[Effect of Bacillus Calmette-Guerin on the expansion of dendritic cells from peripheral blood of pediatric patients with leukemia in vitro].

This study was purposed to investigate the effect of bacillus calmette-guerin (BCG) on the expansion of human dendritic cells (DC) from peripheral blood of pediatric patients with leukemia in vitro. The experiment was divi-ded into two groups: the control and the test group, and the latter group was divided into 3 subgroups: BCG (only BCG), GTI (GM-CSF, TNF-α, IL-4) and GTIB (GM-CSF, TNF-α, IL-4 plus BCG). On day 9 of culture the DCs were counted in each groups, the phenotypes of DC were determined by flow cytometry and these DC were stained with Wright-Giemsa for observation and photography under microscopy. The results showed that the test groups all obtained a certain amount of typical DC; the number of DC in the BCG subgroup is lower than that in the GTI and GTIB subgroups (t=4.20; 6.36, p<0.01); there was no significant difference between the GTI and the GTIB subgroups (t=2.25; p>0.05). The rate of CD1a+ in the BCG subgroup was obviously higher than that in the control group (t=3.04, p<0.05), but was lower than that in the GTI and the GTIB subgroups (t=2.79, 6.41, p<0.05), there was no significant difference between the GTI and the GTIB subgroups (t=0.65, p>0.05). The rate of HLA-DR+, CD83+ in the BCG group was higher than that in the control group (t=4.77, 4.15; p<0.05), but lower than that in the GTI and the GTIB subgroups (t=6.65, 3.19; p<0.05). The rate of HLA-DR+, CD83+ in the GTI subgroup was lower than that in the GTIB subgroup (t=5.64, 2.98; p<0.05). It is concluded that BCG not only promotes the proliferation of DC derived from human peripheral blood of leukemia patients in vitro, but also cooperates with rhGM-CSF, rhTNF-α and rhIL-4 in promoting the maturation of DCs.

[Proliferative inhibition and apoptotic induction effects of crocin on human leukemia HL-60 cells and their mechanisms].

This study was to investigate the proliferative inhibition and apoptosis of human leukemia HL-60 cells induced by crocin and their possible mechanisms. The cell viability was tested by cell counting. The morphology of HL-60 cells was observed by fluorescence microscopy. The MTT assay was used to evaluate the inhibitory effect of crocin on the growth of HL-60 cells. Flow cytometry was used to measure the cell cycle. RT-PCR was used to detect bcl-2 and bax expression. The results indicated that the growth of HL-60 cells was inhibited remarkably in the dose and time dependent way. When the crocin concentration was higher than 5 mg/ml, the percentage of apoptotic HL-60 cells was not increased, on the contrary this percentage decreased, the cells manifested necrosis. Flow cytometry profiles revealed that cells were blocked in G0/G1 phase, the cell proliferation was inhibited obviously at 5 mg/ml. RT-PCR detection revealed that the expression of bcl-2 was down-regulated strikingly and bax was up-regulated. It is concluded that the crocin can inhibit the proliferation of HL-60 cells effectively, and block cells in G0/G1 phase. The mechanisms by which crocin induced apoptosis in HL-60 cells may be related to the inhibition of bcl-2 and activation of bax.

[Influence of Interferon alpha-2b on proliferation inhibition and apoptosis induction in HL-60 cells].

To investigate the effects of interferon alpha-2b on proliferation and apoptosis in HL-60 cells, HL-60 cells were cultured in different concentrations of IFN alpha-2b. The morphologic changes were observed by Wright's and acridine orange (AO) and ethidium bromide (EB) staining respectively. Inhibition of proliferation was detected by MTT. Expression of CD13(+) was checked by indirect fluoroimmunoassay. The results showed that apoptosis rate of HL-60 cells assayed by the above-mentioned two methods was (51 +/- 2)% and (78 +/- 3)% respectively and OD(570) values of proliferation inhibited were 1.8 +/- 0.1 and 1.0 +/- 0.1 respectively when the concentrations of the IFN(alpha-2b) were 500 and 10,000 U/ml in culture for 48 hours. Morphology and count of CD13(+) cells were changed. CD13(+) cell expression rate was (62 +/- 2)% and (30 +/- 3)% respectively when the concentrations of the IFN(alpha-2b) were 500 and 10,000 U/ml in culture for 48 hours. It is concluded that IFN(alpha-2b) can enhance the apoptosis of HL-60 cells, inhibit their proliferation, promote their maturation and differentiation.

[Effect of nimodipine on mechanisms of HL-60 cell apoptosis induced by cytarabine].

The aim was to study the mechanisms of HL-60 cell apoptosis induced by nimodipine (NMDP) and cytarabine (Ara-C). The DNA fragment was detected by agarose gel electrophoresis. The expressions of bcl-2 and bax gene proteins related with apoptosis were investigated by immunohistochemistry. The results showed that HL-60 cell apoptosis rate had been increasing in the experimental groups compared with the control group since culturing 8 hours. The expression of Bcl-2 protein was lower and the expression of Bax protein was higher in the experimental groups than that in the control group, while ratio of bcl-2/bax was lower in the experimental groups than that in the control group. It is concluded that NMDP and Ara-C induce apoptosis of HL-60 cells, and the mechanism of apoptosis induced by them may down-regulate the expression of bcl-2 gene and up-regulate the expression of bax gene. The mechanism of HL-60 cell apoptosis induced by Ara-C and NMDP is probably associated with the down-regulation of Bcl-2 protein expression.