Cardiac optogenetics: a novel approach to cardiovascular disease therapy.

Optogenetics is a cell-type specific and high spatial-temporal resolution method that combines genetic encoding of light-sensitive proteins and optical manipulation techniques. Optogenetics technology provides a novel approach for research on cardiac arrhythmia treatment, including pacing, recovering the conduction system, and achieving cardiac resynchronization with precise and low-energy optical control. Photosensitive proteins, which usually act as ion channels, pumps, or receptors, are delivered to target cells, where they respond to light pulses of specific wavelengths, evoke transient flows of transmembrane ion currents, and induce signal transmission. With the development of gene technology, the in vivo efficiency of optogenetics in cardiology has been trialed, and in vitro experiments have been performed to test its potential in cardiac electrophysiology. Challenges for applying optogenetics in large animals and humans include the effectiveness, safety, and long-term expression of photosensitive proteins, unscattered and unattenuated exogenous light stimulation, and the need for implantable miniature light stimulators. Photosensitive proteins, genetic engineering technology, and light equipment are essential for experiments in cardiac optogenetics. Optogenetics may provide an alternative method for evaluating the mechanism of cardiac arrhythmias, testing hypotheses, and treating cardiovascular diseases.

Apoptosis in human myelodysplastic syndrome CD34+ cells is modulated by the upregulation of TLRs and histone H4 acetylation via a ß-arrestin 1 dependent mechanism.

Excessive apoptosis of hematopoietic precursors in the bone marrow underlies the ineffective hematopoiesis characteristic of myelodysplastic syndrome (MDS). Toll-like receptor (TLR) signaling is abnormally activated in MDS and may be involved in excessive programmed cell death in the pathogenesis of MDS. TLRs expression and global histone H3/H4 acetylation were analyzed in bone marrow (BM) CD34+ cells from 20 lower-risk and 20 higher-risk MDS patients and 10 healthy controls. Apoptosis of BM CD34+ cells was examined by flow cytometry, and its correlation to histone acetylation and the expression of TLR2 and ß-arrestin1 (ß-arr1), measured by enzyme-linked immunosorbent assay and qRT-PCR, was assessed. TLR1, TLR2 and TLR6 expression and H4 acetylation levels were higher in lower-risk MDS patients than in higher-risk MDS patients or controls, and TLR2 expression and H4 acetylation levels were positively correlated with an increased rate of apoptosis. Lower-risk MDS was associated with increased ß-arr1 expression and histone acetyltransferase p300 activity. In in vitro-cultured primary normal and lower-risk MDS CD34+ cells, TLR2 activation-induced apoptosis was mediated by the upregulation of ß-arr1 leading to the recruitment of p300 and increased histone H4 acetylation. The nuclear accumulation of ßarr1 following TLR2 activation promote H4 acetylation at specific target gene promoters and may thus affect transcription of target genes in BM CD34+ cells. The mechanisms underlying the deregulation of TLR2 and increased apoptosis in MDS may involve the ß-arr1 mediated recruitment of p300 leading to increased levels of histone H4 acetylation.

[Influence of Ginkgo biloba extract on proliferation of ACC-2 cell, Survivin and TIP30 gene expression in adenoid cystic carcinoma of lacrimal gland].

Exploring the influence of extract of Ginkgo biloba (EGB) on the proliferation, apoptosis of ACC-2 cell in lacrimal adenoid cystic carcinoma and analyzing the influence of EGB on the gene expression of Survivin and TIP30 based on the levels of the gene and protein. ACC-2 cell in human with ACC of lacrimal gland disposed by EGB of different concentration was in vitro cultured. MTT method was used for cell proliferation detection. Annexin V/PI double-staining flow cytometer was used to detect cell apoptosis and cell cycle. Survivin and TIP30 gene expression together with protein expression were analyzed by RT-PCR and Western blotting. And it is indicated that EGB has inhibitory effect on the proliferation of ACC-2 cell in vitro. Furthermore, the dose-effect relationship was significant. Compared with the control group, it had statistical difference (P <0.01). The inhibitory concentration 50% (ICso) is 88 mg . L-1. By flow cytometer examination, it was indicated that EGB can gradually increase ACC-2 cell in G0-G1 stage and decrease it in G2-M and S stage. With the increase of dose, the apoptosis rate of ACC-2 cell obviously increased (P <0.05 or P <0.01). Both of the expression results of RT-PCR and Western hybrid proteins have showed that the concentration of EGB increased, it could be seen a significant decrease in Survivin gene expression (P <0.01). Meanwhile, the TIP30 gene expression got a significant increase. Therefore, EGB can effectively inhibit ACC-2 cell Survivin gene expression in human with adenoid cysistic carcinoma of larcrimal gland as well as promoting TIP30 gene expression, inducing the ACC-2 cell apoptosis and inhibiting tumor cell proliferation, which provided a certain theoretical and experimental basis for the application of Chinese herbal medicinal ingredient in the treatment of tumors.

A circRNA ceRNA network involved in cognitive dysfunction after chronic cerebral hypoperfusion.

Chronic Cerebral Hypoperfusion (CCH) is associated with cognitive dysfunction, the underlying mechanisms of which remain elusive, hindering the development of effective therapeutic approaches. In this study, we employed an established CCH animal model to delve into neuropathological alterations like oxidative stress, inflammation, neurotransmitter synthesis deficits, and other morphological alterations. Our findings revealed that while the number of neurons remained unchanged, there was a significant reduction in neuronal fibers post-CCH, as evidenced by microtubule-associated protein 2 (MAP2) staining. Moreover, myelin basic protein (MBP) staining showed exacerbated demyelination of neuronal fibers. Furthermore, we observed increased neuroinflammation, proliferation, and activation of astrocytes and microglia, as well as synaptic loss and microglial-mediated synapse engulfment post-CCH. Utilizing RNA sequencing, differential expression analysis displayed alterations in both mRNAs and circRNAs. Following gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, both showed significant enrichment in immunological and inflammation-related terms and pathways. Importantly, the differentially expressed circular RNAs (DE circRNAs) exhibited a notable coexpression pattern with DE mRNAs. The ternary circRNA-miRNA-mRNA competing endogenous RNAs (ceRNA) network was constructed, and subsequent analysis reiterated the significance of neuroimmunological and neuroinflammatory dysfunction in CCH-induced neuropathological changes and cognitive dysfunction. This study underscores the potential role of circRNAs in these processes, suggesting them as promising therapeutic targets to mitigate the detrimental effects of CCH.

[Research Progress of Thalidomide and Its Derivatives in Treatment of Myelodysplastic Syndrome--Review].

Thalidomide and its derivatives have been used in the treatment of myelodysplastic syndrome (MDS) because of their anti-angiogenic and immunomodulatory effects. In recent years, some studies have found that thalidomide and its derivatives not only showed significant efficacy in lower-risk MDS patients with del (5q), but also showed advantages in non-del (5q) MDS patients. In addition, the discovery of its molecular targets and new substrates makes it possible to develop a new generation of immunomodulatory drugs (IMiDs) and to design IMiDs-based proteolysis-targeting chimeras. In this review, the new progress in mechanism and clinical application of thalidomide and its derivatives were summarized briefly, so as to provide a more scientific, reasonable and effective scheme to the treatment of MDS.

[Advances in the Action Mechanism of Classical Pathways IKKα and IKKß in Hematological Tumors and Drug Therapy Blocking Their Effect--Review].

In recent years, it is found that the classical IKKα and IKKß pathway were closely relates with hematological tumors, except the classical pathogenesis, moreover the classical IKKß pathway is deeply studied. The studies indicated that the IKKßis activated to phosphorylate the NF-κB through multiple cascades under the effect of extracellular IL-6, TNF-α and other stimulating factors. At the cellular level, the classical IKKßcan promote the tumor cell survival and proliferation, reduce the cell apoptosis, and promote the angiogenesis and cell transfer. Although the classical IKKα plays a role in regulating IKKß activity, but its role in non-classical pathway is more prominent. This review briefly summarizes the latest advance of researches on the pathogenesis of hematological malignancies in term of IKKα and IKKßpathway, so as to provide the theoretic basis for deeply understanding and studying the pathogenesis of hematologic tumors. At present, blocking the classical IKKα and IKKß pathway has become a new target for treatment of hematological tumors, moreover, some specific inhibitor for IKKα and IKKßpathway have been developed, for example, LY2409881, BMS 345541 and so on. Most of these drugs are in clinical trials and display some good anti-tumor effects.

Anti-tumor and Phenotypic Regulation Effect of Matrine on Dendritic Cells through Regulating TLRs Pathway.

OBJECTIVE: To investigate the effects of matrine on antigen presentation of dendritic cells (DCs), and to explore the pharmacological mechanism of matrine on anti-tumor effect. METHODS: Different concentrations (0, 1, 2, 4, 8 and 16 µ g/mL) of matrine were co-cultured with DCs, the harvested DCs were co-cultured with antigens of Lewis lung cancer (LLC) cells, and then DCs and T cells were co-cultured to produce DCs-activated killer (DAK) cells, which have significant tumor-killing activity. The expression of cytokines, mRNA and protein of toll-like receptors (TLRs) in DCs were detected by enzyme linked immunosobent assay, polymerase chain reaction and Western blot, respectively. And the killing effect of DAK were measured by MTT assay. RESULTS: Matrine significantly increased the mRNA expression of TLR7, TLR8, myeloid differentiation factor 88 (MyD88), tumor necrosis factor receptor-associated factor 6 (TRAF-6) and I κ B kinase (IKK), as well as the protein expression of TLR7 and TLR8, and up-regulated the levels of interleukin-12 (IL-12), IL-6 and tumor necrosis factor-α (TNF-α), meanwhile, it also increased the expressions of MHC-II, CD54, CD80 and CD86 in DCs. DCs-activated effector T cells had significant tumor-killing activity. When the concentration of matrine was more than 4 µg/mL, all indices had significant difference (P<0.01 or P<0.05). CONCLUSION: Matrine plays an anti-tumor role by regulating TLRs signal transduction pathway, promoting the secretion of inflammatory cytokines and enhancing immune function.

[Influence of Shengxue mixture on the expression of T-bet/GATA-3, their relevant signal transduction molecules, cytokines, and Th1/Th2 balance in patients with chronic aplastic anemia].

OBJECTIVE: To study the actions of transcription factors, T-bet and GATA-3, and their relevant signal transduction pathways on the immune-related pathogenesis with chronic aplastic anemia (CAA), and to investigate the immunological regulation mechanism of Shengxue Mixture (SXM) in regulating levels of Th cell imbalance, transcriptional factor and relevant signal pathways. METHODS: All CAA patients selected from Yueyang Hospital of Shanghai University of traditional Chinese medicine were equally randomized into the treated group and the control group, 20 patients in each group, and 20 healthy persons were selected as normal group, the former was treated with SXM according to patients' syndrome patterns, namely, SXM-1 was given to patients of Pi-Shen yang-deficiency pattern, and SXM-2 to those of Pi-Shen yin-deficiency pattern. Patients in the control group were treated with cyclosporin A (CsA). The mRNA expressions of T-bet, GATA-3, signal transducers and activators of transcription 4 (STAT4) and 6 (STAT6) in peripheral blood mononuclear cell (PBMNC) of patients were determined using real-time fluorescent quantitation polymerase chain reaction before and after treatment, meantime, the Th1/Th2 proportion in peripheral blood, and levels of IFN-gamma, IL-12 and IL-4 in PBMNC-cultured supernatant were detected by flow cytometry and enzyme linked immunosorbent assay. RESULTS: The mRNA expressions of PBMNC T-bet and STAT4, ratios of T-bet/GATA-3, Th1 proportion and Th1/Th2 ratio, levels of IFN-gamma and IL-12 in PBMNC-cultured supernatant were all significantly higher in CAA patients than in healthy controls (P < 0.01), which were lowered after treatment but didn't reach the normal range (all P < 0.01), excepting for IL-12 level. Comparisons of the changes between the two treated groups showed insignificant difference (P > 0.05). While the difference between patients and healthy persons in terms of GATA-3, STAT6, Th2 proportion, and IL-4 were insignificant (P > 0.05), either before or after treatment. CONCLUSIONS: Abnormal activation of IFN-gamma/T-bet and IL-12/ STAT4 pathways, as well as Th1/Th2 balance deviating to Th1 excursion play vital roles in the immunological pathogenesis of CAA. SXM and CsA could lower the aforesaid abnormal activation and correct Th1 hyper-polarization, so as to alleviate the over-activated cell-mediated immunity to eliminate hematopoietic depression in CAA patients.

[Research Progress on Immune Mechanism of Immune Thrombocytopenia--Review].

Abstract　　Immune thrombocytopenia (ITP) is an acquired autoimmune hemorrhagic disease, although the ITP pathogenesis is completely unknown, but in terms of the current view, the immune tolerance is main reason for the onset of ITP. In recent years, more and more immune cell subsets, cytokines and the new approacher were found to be closely related with the ITP, such as saliva acid, B cell activating factor, dysfunction of regulatory B cells and Th1/Th2 balance drift, CD4+ CD25+ T cell function defect, IL-23/Th17 pathway regulation, etc., In this paper, the latest research progress on the immune pathogenesis of ITP are reviewed, so as to provide theoretical basis and research direction for further understanding the pathogenesis of ITP.

[Increased expression of transient receptor potential melastatin 7 in mouse cardiac fibroblasts post myocardial infarction].

OBJECTIVE: To observe transient receptor potential melastatin 7-like (TRPM7L) expression changes post myocardial infarction (MI) in mouse cardiac fibroblast (CF). METHODS: TRPM7 expression and Ca2+ influx in CF from MI and control mice were quantified by mRNA RT-PCR and whole cell patch clamp technique. RESULTS: (1) TRPM7 expression was significantly upregulated post MI and Ca2+ influx of CF were significantly increased post MI [(7.4 +/- 0.7) pA/pF vs. (16.2 +/- 1.7) pA/pF, P < 0.01] and Ca2+ influx of CF increased 3-fold under lower pH condition; (2) These effects could be blocked by knock-out TRPM7 gene with SiRNA. CONCLUSION: TRPM7L upregulation post MI and under lower pH condition are responsible for increased Ca2+ influx in CF.

[Effects of Yisui Jiedu Recipe on JAK2-STAT5 signal transduction pathway in bone marrow hematopoietic cells from patients with myelodysplastic syndrome-refractory anemia].

OBJECTIVE: To investigate the effect of Yisui Jiedu Recipe (YSJDR), a compound traditional Chinese herbal medicine, on cytokines and their corresponding just another kinase 2-signal transducers and activators of transcription 5 (JAK2-STAT5) signal transduction pathway in bone marrow hematopoietic cells from patients with myelodysplastic syndrome-refractory anemia (MDS-RA). METHODS: Fluorogenic quantitative polymerase chain reaction (FQ-PCR) method was established to detect the levels of JAK2, STAT5 and Bcl-xL mRNA expressions, and JAK2-STAT5 signal transduction pathway was activated by granulocyte-macrophage-colony stimulating factor (GM-CSF) in cultured bone marrow hematopoietic cells from 10 patients with MDS-RA. The levels of interleukin-2 (IL-2), interleukin-3 (IL-3), gamma-interferon (gamma-INF) and tumor necrosis factor-alpha (TNF-alpha) in the cultural supernatant of untreated control, AG490-treated and YSJDF-treated cells were measured by enzyme-linked immunosorbent assay. RESULTS: The levels of IL-2 and TNF-alpha in YSJDR-treated group were significantly lower than those in untreated control group and AG490-treated group (P<0.01, P<0.05), and IL-3 level in YSJDP-treated group was remarkably higher than that in the other two groups (P<0.01). There were no significant differences in the levels of IL-2 and IL-3 between AG490-treated group and untreated control group (P>0.05), while the TNF-alpha level in AG490-treated group was decreased obviously as compared with the untreated control group (P<0.01). There was no significant difference in gamma-INF level between YSJDR-treated group and AG490-treated group (P>0.05), while TNF-alpha level in the two groups were significantly lower than that in the untreated control group (P<0.01). The expressions of JAK2, STAT5 and Bcl-xL mRNAs were significantly down-regulated in the YSJDR-treated and the AG490-treated groups as compared with those in the untreated control group (P<0.05, P<0.01), while there were no differences in the expressions of JAK2, STAT5 and Bcl-xL mRNAs between YSJDR-treated group and AG490-treated group. CONCLUSION: YSJDR can modulate cytokine level in bone marrow hematopoietic cells of MDS-RA, suppress JAK2-STAT5 signal transduction, and inhibit the Bcl-xL mRNA expression.

Malignancy as a comorbidity in rheumatic diseases: a retrospective hospital-based study.

Patients with Rheumatic diseases (RDs) are at an increased risk of malignancies compared with the general population. The aim of this study was to examine the relative frequency of several cancers in a single homogeneous cohort of patients with different RDs. Patients diagnosed with rheumatoid arthritis (RA), Ankylosing spondylitis (AS), Sjögren's syndrome (SS), systemic lupus erythematosus (SLE), systemic sclerosis (SSc), dermatomyositis (DM), or polymyositis were included. Out of 3982 adult residents admitted to the division of rheumatology, 61 malignancies were observed. The 2009 National Central Cancer Registry (NCCR) of China served as the reference for calculating standardized ratio (SR). The malignancy frequency had no difference between RDs with malignancy and the general population. Patients with SS and DM/PM showed an increased risk of non-Hodgkin's lymphoma (SR for SS patients = 9.709, 95% confidence interval (CI) = 4.602 to 17.916; SR for DM/PM = 35.714, 95% CI = 25.001 to 49.527). Patients with DM/PM and SSc showed an increased risk of lung cancer (SR for DM/PM = 10.638, 95% CI = 5.245 to 19.131; SR for SSc patients = 7.752, 95% CI = 3.295 to 15.309). Patients with SS and DM/PM showed an increased risk of ovary cancer (SR for SS patients = 8.177, 95% CI = 3.566 to 15.888; SR for DM/PM = 32.258, 95% CI = 22.126 to 45.490). Patients with SLE showed an increased risk of cervix cancer (SR for AS patients = 6.897, 95% CI = 2.748 to 14.144). Patients with AS showed an increased risk of pancreas cancer (SR for AS patients = 7.576, 95% CI = 2.181 to 15. 071). Different RDs have an increased risk of particular cancers. Among hematologic cancers, the risk of non-Hodgkin's lymphoma was higher than general population. Among solid tumors, the risk of cancers of the lung, ovary, cervix, and pancreas was higher than general population.

Antibiotic resistance and molecular characterization of diarrheagenic Escherichia coli and non-typhoidal Salmonella strains isolated from infections in Southwest China.

BACKGROUND: Bacterial diarrhea is one of the most common causes for medical consultations, mortality and morbidity in the world. Diarrheagenic Escherichia coli (DEC) and non-typhoidal Salmonella (NTS) are major intestinal pathogens in developing countries, and the indiscriminate use of antibiotics has greatly contributed to resistant strains. Hence, the aim of the present study is to identify the antimicrobial resistance patterns and the molecular characteristics of DEC and NTS in southwest, China. METHODS: 1121 diarrheal patients and 319 non-diarrheal subjects across all age groups were recruited from four sentinel hospitals from June 2014 to July 2015 in Kunming City, Yunnan Province. Each stool specimen was collected to detect DEC and NTS with standard microbiological and molecular methods. Antimicrobial resistance testing was performed with the Kirby-Bauer disk diffusion method, and the standards for antimicrobial susceptibility testing complied with the Clinical and Laboratory Standards Institute (CLSI). Molecular characterization of strains was carried out using pulsed-field gel electrophoresis (PFGE). A structured questionnaire was used to record basic epidemiological data (e.g. sex, age, residence, season, etc.). Data were analyzed using Chi-square or Fisher's exact test. RESULTS: DEC was detected in 127 (11.33%) diarrhea cases and 9 (2.82%) non-diarrheal cases (χ2 = 20.69, P < 0.001, OR = 4.36, 95% CI: 2.19-8.65), and the prevalence of NTS isolated from diarrhea cases was higher than that of non-diarrheal cases across all age groups (n = 42, 3.75%, n = 1, 0.31%, χ2 = 10.10, P = 0.002, OR = 12.38, 95% CI: 1.70-90.29). The rates of resistance to ten antibiotics of DEC and NTS showed significant differences (χ 2 = 386.77, P < 0.001; χ2 = 191.16, P < 0.001). The rates of resistance to Amoxicillin and Clavulafiate (AMC), Cephalothin (CEP), Gentamicin (GEN) and Sulfamethoxazole-Trimethoprim (SXT) of DEC isolated from diarrhea cases were higher than those of NTS isolated from diarrhea patients (37.01% vs 14.29%, χ2 = 7.57, P = 0.006; 29.92% vs 11.90%, χ2 = 5.40, P = 0.02; 37.01% vs 11.90%, χ2 = 5.80, P = 0.016; 62.20% vs 26.19%, χ2 = 16.44, P < 0.001; respectively). Ciprofloxacin (CIP) was the most sensitive antibiotic for DEC and NTS strains isolated from diarrhea cases. Resistance rates of DEC isolates from cases and controls to more than three kinds antimicrobials (multidrug resistance, MDR) showed no significant differences (81.10% vs 88.89%, P = 0.33). Pulsotype patterns of DEC strains were highly diverse; however, the pulsotype pattern of NTS strains was closely related to the serotype. The pattern of S. enteritidis was highly similar, but the S. enterica Typhimurium strain was discrete. CONCLUSIONS: Antibiotic resistance of Enterobacteriaceae is of great concern. The societal effects of antibiotic use justify strict monitoring to combat increases in antimicrobial resistance. Molecular epidemiology and systematic epidemiological investigation can provide accurate evidence for tracking the infection source.

[Role of MG132, a proteasome inhibitor, in inducing expression of costimulatory molecules CD86 in leukemic cells and its effect on allogeneic mixed lymphocyte reaction].

OBJECTIVE: To investigate the role of proteasome inhibitors MG132 in the inducing the expression of the costimulatory molecules CD80 and CD86 in leukemia cells and its effect on allogeneic mixed lymphocyte reaction. METHODS: Acute myelocytic leukemia cells of the line HL-60 and chronic myelocytic leukemia cells of the line K562 were cultured. 7-AAD staining and flow cytometry (FC) were used to examine the viability of the cells. MG132, a proteasome inhibitor, of the concentrations of 2 or 3 micromol/L was added into the culture fluid of HL-60 cells for 24 h and 48 h respectively and then annexin V/7-AAD staining and FC were used to detect the apoptosis of the cells. HL-60 and K562 cells treated with 1 micromol/L MG132 for 24 h and 48 h respectively, anti-CD80 and anti-CD86 antibodies were added, then FC was used to detect the expression of CD80 and CD86. The mRNA expression of CD86 in the HL-60 cells treated with 1 micromol/L MG132 was examined by RT-PCR. HL-60 and K562 cells were treated by 1 micromol/L MG132 for 48 h and then underwent irradiation of 75 Gy Co-60 to kill the cells with their antigenicity preserved. Peripheral blood mononuclear cells (PBMNC) of healthy volunteers, as reactive cells, were isolated and inoculated into the Co-60 treated HL-60 and K532 cells of different concentrations, as stimulating cells, for 5 d, CCK-8, a new agent to detect the cell viability, was added for 4 h, and then the A value of absorbance was measured at the wave length of 450 nm of enzyme labeling instrument. Control groups were set up for all tests. RESULTS: The cell viability rates of the HL-60 cell treated with 1 micromol/L MG132 for 24 h and 48 h were 92.95% and 85.87% respectively. The apoptotic rats of the HL-60 cells treated with MG132 were increased dose- and time-dependently. Before MG132 treatment K562 cells did not express CD86, and the CD86 expression of the HL-60 cells was up-regulated time-dependently (all P < 0.01). The mRNA expression of CD86 in the HL-60 treated with MG132 was up-regulated time-dependently (P < 0.01). CKK8 test showed that the proliferation level of PBMNC gradually increased along with the concentration of HL-60 cells treated with MG132 and reached its peak when the concentration of the HL-60 cells was 1 x 10(5) (P < 0.01). No remarkable proliferation of PBMNC was seen in the K562 groups no matter if the HL-60 cells had been treated with MG132. CONCLUSION: MG132 induces the expression of costimulatory molecule CD86 in the HL-60 cells, thus improving the proliferation of PBMNC.

[Effects of Jianpi Bushen Huoxue recipe on hematopoietic cell apoptosis in a mouse model of aplastic anemia].

OBJECTIVE: Adopting methods of cell culture to explore the effects and mechanisms of Jianpi Bushen Huoxue Prescription (JPBSHXP), a traditional Chinese compound herbal medicine for strengthening spleen, reinforcing kidney and activating blood circulation, in inhibiting hematopoietic cells apoptosis in a mouse model of aplastic anemia (AA). METHODS: Blood serum of AA mice was made from an AA mouse model. Blood serums containing different traditional Chinese compound herbal medicine were made from rats after intragastric administration of JPBSHXP and its related decoctions, respectively. Bone marrow cells of normal mice were incubated by these blood serums for 24 hours, respectively. The apoptosis of the bone marrow cells were assayed by flow cytometry and transmission electron microscopy (TEM). RESULTS: It was indicated that the bone marrow cells of normal mice incubated with blood serum of AA mice displayed typical apoptosis. The apoptosis rates of bone marrow cells of the AA mice incubated by blood serum containing different traditional Chinese herbal medicine were decreased. The effect of Bushen (reinforcing kidey) Recipe was better than Jianpi (strengthening spleen) Recipe and Huoxue (activating blood circulation) Recipe, while the effect of JPBSHXP was the best. TEM results showed that the effect of Bushen Recipe was better than that of the Jianpi Recipe and the Huoxue Recipe, while the effect of JPBSHXP was the best. CONCLUSION: JPBSHXP and its related decoctions can significantly decrease the apoptosis rate of bone marrow mononuclear cells of the AA mice. It is inferred that JPBSHXP can promote bone marrow hematogenesis.

Effects of arsenic disulfide on apoptosis, histone acetylation, toll like receptor 2 activation, and erythropoiesis in bone marrow mononuclear cells of myelodysplastic syndromes patients in vitro.

OBJECTIVE: As the main component of traditional Chinese medicine realgar, arsenic disulfide (As2S2) is widely used in treating myelodysplastic syndromes (MDS). The goal of the current study is to assess the effects of As2S2 on bone marrow mononuclear cells (BMMNC) of MDS. METHODS: BMMNCs were obtained from 10 lower risk MDS patients, 5 higher risk MDS patients, and 3 healthy controls. Then, the cells were treated with As2S2 for 48h, using vorinostat (also known as SAHA) as control. Cell proliferation and apoptosis were detected. mRNA and protein levels of histone deacetylase-1 (HDAC1), Toll-like receptor 2 (TLR2), and erythroid transcription factor (GATA-1) were detected by quantitative real-time PCR and western blot analysis. RESULTS: After As2S2 treatment in concentrations ranging from 3.125 to 100µmol/L, cell proliferation was inhibited in both lower risk and higher risk MDS. Fifty percent inhibitory concentrations were 24.4µmol/L and 23.6µmol/L, respectively, for lower and higher risk MDS. Apoptotic cells significantly increased in both types of MDS. mRNA and protein levels of HDAC1 and TLR2 were reduced, whereas GATA-1 was increased in both types of MDS. CONCLUSIONS: As2S2 could inhibit cell proliferation and induce apoptosis through histone acetylation modulation in MDS. Similar to SAHA, As2S2 could reduce TLR2 activation and increase GATA-1 expression. Current data suggest epigenetic and immunological alternations are involved in therapeutic mechanisms of realgar in the treatment of MDS.

[Changes in T-cell receptor repertoire in aplastic anemia and effects of Shengxue Mixture].

OBJECTIVE: To explore the immune pathogenesis of aplastic anemia (AA) and the therapeutic effects of Shengxue Mixture (SM) through the gene expressions of subfamilies of T-cell receptor variable region beta (TCR Vbeta) using immunologic and molecular biologic technology. METHODS: Gene expressions of TCR Vbeta sub-families in peripheral blood mononuclear cells from 20 AA patients were detected before and after treatment with SM using RT-PCR and gene scanning method. RESULTS: TCR Vbeta gene repertoire of the 24 subfamily genes deviated in AA patients, and the oligoclonal gene expressions increased obviously compared with those in healthy people (P < 0.01), including Vbeta2, 5, 6, 15, 16, 22, and 23 were found in 30%-50% AA patients, and Vbeta8, 21 were in more than 50% patients. These oligoclonal genes reduced significantly after treatment with SM compared with those before treatment (P < 0.05). CONCLUSION: Multiple TCR Vbeta subfamilies of clonal proliferation participate in the pathogenesis of AA. SM can rectify the deviation of TCR Vbeta gene repertoire, reduce the abnormal clonal proliferation of T cells, thus to alleviate the immune injury to hematopoietic tissue, and thus to benefit the recovery of hematopoiesis of bone marrow.

Qualitative analysis on a proposal for key specific medical treatment for aplastic anemia.

OBJECTIVE: By reviewing the medical treatments for aplastic anemia (AA, Suilao Disease), which is the important research interest of Collaborative Group, Key Department of Blood Disease, State Administration of Chinese medicine, the consensus on the diseases have been reached among the different units of the collaborative group. METHODS: Using qualitative analysis, we determined the characteristics, location and pathogenesis of Suilao disease. We discovered the ways of traditional Chinese medical treatment in curing Suilao disease. RESULTS: Acute AA (acute suilao) and chronic AA (chronic suilao) diseases require different treatment. Acute AA requires 3 phrases of treatments, which are "cold", "warm" and "hot". However, chronic AA requires a dialectic treatment, which involves reinforcement of the Shen (Kidney). Suitable Chinese medical treatments for curing Suilao disease were discussed and reached a consensus. CONCLUSION: It is concluded that a summarized therapy approved by many experts could be widely used.

Impact of co-infections with enteric pathogens on children suffering from acute diarrhea in southwest China.

BACKGROUND: Acute diarrhea is a global health problem, resulting in high morbidity and mortality in children. It has been suggested that enteric pathogen co-infections play an important role in gastroenteritis, but most research efforts have only focused on a small range of species belonging to a few pathogen groups. This study aimed to assess the impact of co-infections with a broad range of enteric pathogens on children aged below five years who suffer from acute diarrhea in southwest China. METHOD: A total of 1020 subjects (850 diarrhea cases and 170 healthy controls) were selected from four sentinel hospitals in Kunming, Yunnan province, southwest China, from June 2014 to July 2015. Stool specimens were collected to detect five virus (rotavirus group A, RVA; norovirus, NoV; Sapovirus, SaV; astrovirus, As; and adenovirus, Ad), seven bacterial (diarrheagenic Escherichia coli, DEC; non-typhoidal Salmonella, NTS; Shigella spp.; Vibrio cholera; Vibrio parahaemolyticus; Aeromonas spp.; and Plesiomonas spp.), and three protozoan (Cryptosporidium spp., Giardia lamblia, and Blastocystis hominis, B. hominis) species using standard microbiologic and molecular methods. Data were analyzed using the partial least square regression technique and chi-square test. RESULTS: At least one enteric pathogen was detected in 46.7 % (n = 397) of acute gastroenteritis cases and 13.5 % (n = 23) of healthy controls (χ(2) = 64.4, P < 0.05). Single infection with RVA was associated with acute diarrhea (26.5 % vs. 5.8 %, P < 0.05). The prevalence of a single infection with B. hominis in diarrhea cases was higher than in healthy controls (3.1 % vs. 0.5 %, OR = 4.7, 95 % CI: 1.01-112.0). Single infection with NoV GII was not associated with diarrhea (4.4 % vs. 3.5 %, OR = 1.2, 95 % CI: 0.5-3.3). Single infections with bacterial species were not observed. The prevalence of co-infections with two enteric pathogens in diarrhea cases was higher than in asymptomatic children (20.1 % vs. 5.3 %, P < 0.05). RVA-NoV GII was the most common co-infection in symptomatic children (4.4 %), with it aggravating the severity of diarrhea. CONCLUSIONS: Although it is clear that RVA has an overwhelming impact on diarrhea illnesses in children, co-infection with other enteric pathogens appears to also aggravate diarrhea severity. These findings should serve as evidence for public health services when planning and developing intervention programs.