MRI findings of central nervous system involvement in children with haemophagocytic lymphohistiocytosis: correlation with clinical biochemical tests.

AIM: To investigate the brain magnetic resonance imaging (MRI) features of children with haemophagocytic lymphohistiocytosis (HLH) with central nervous system (CNS) involvement, and to investigate the correlation with clinical biochemical tests. MATERIAL AND METHODS: Clinical and MRI data were collected from 118 children with HLH-CNS between January 2012 and June 2019. Patients were grouped according to their MRI findings, and statistical methods were used to test for correlations between the MRI findings and biochemical variables. RESULTS: Patients were divided into three groups, including normal appearance (Group 1, 17/118), diffuse parenchymal volume loss (Group 2, 44/118), and brain parenchyma lesions (Group 3, 57/118) containing three subtypes of brain lesions and HLH-CNS complications. Comparing biochemical values among the three groups revealed a significant difference for all values (p<0.05), except for cell counts in the cerebrospinal fluid (CSF). A pairwise comparison further showed significant inter-group differences for most of the variables. Spearman's rank correlation coefficient also demonstrated that CSF cell counts (r=0.193, p=0.036), CSF microprotein content (r=0.379, p<0.001), serum aspartate aminotransferase (AST; r=0.521, p<0.001), serum lactate dehydrogenase (LDH; r=0.514, p<0.001) and activated partial thromboplastin time (APTT; r=0.326, p<0.001) correlated positively with the MRI groups, while platelet count (PLT; r=-0.633, p<0.001) and plasma fibrinogen (FIB; r=-0.258, p=0.005) correlated negatively. CONCLUSION: Classification of brain MRI findings of HLH-CNS correlates well with the results of several key biochemical tests. Brain MRI is a promising method to elucidate illness severity and clinical outcomes.

Hydroxytyrosol suppresses LPS-induced intrahepatic inflammatory responses via inhibition of ERK signaling pathway activation in acute liver injury.

OBJECTIVE: Acute liver injury (ALI) leads to inflammatory response and tissue damage. Inflammatory activation of infiltrative macrophages plays a critical role in liver histology destruction and dysfunction. Hydroxytyrosol (3,4-dihydroxyphenil-ethanol, HT), one of the polyphenols extracted from extra virgin olive oil, currently acts as a treatment for neuroinflammatory responses, but its effect on ALI is elusive. The present study aims to examine the mechanism of HT in macrophages inflammation and evaluate treatment effect of HT on ALI. MATERIALS AND METHODS: In vitro, the expressions of type M1/M2 macrophages biomarkers (CD11c/CD206) and cytokines (TNF-α, IL-1ß, IL-6, IL-10, IL-4) following lipopolysaccharide (LPS) stimulation and HT administration were detected using immunofluorescence, quantitative real-time polymerase chain reaction (qRT-PCR), and enzyme-linked immunosorbent assay (ELISA). Mechanically, HT was used to treat cells and phosphorylation level of extracellular-signal-regulated kinase 1/2 (ERK1/2) protein in cells was analyzed using Western blotting. In murine acute liver injury, inflammatory cytokines and liver injury degree were exhibited by qRT-PCR, IHC and HE staining. Furthermore, hepatic function was exhibited via hepatic metabolic enzymes (ALT/AST) and total bilirubin (TBil) in serum. RESULTS: It was demonstrated that HT treatment attenuated M1 macrophages and increased M2 macrophages after LPS stimulation. Furthermore, the pro-inflammatory cytokine level was descended, while an-inflammatory cytokine was increased via HT suppressing ERK pathway in macrophages. In vivo, HT reduced inflammatory level and mitigated hepatic histological injury, thus ameliorating liver function after acute liver injury. CONCLUSIONS: HT exerts a hepatoprotective and anti-inflammation effect on acute liver injury, which restrains inflammation by inhibiting ERK pathway and regulating macrophages polarization. Moreover, HT prevents liver tissues from inflammatory injury. Therefore, HT serves as a potential implication to treat ALI through modulating inflammation of macrophages.

[The effect of arthroscopic synovectomy on refractory knee arthritis with popliteal cyst in 153 patients].

Objective: To investigate the efficacy of arthroscopic synovectomy on refractory knee arthritis complicated with popliteal cyst. Methods: Patients diagnosed as rheumatoid arthritis (RA) or spondyloarthritis (SpA) with refractory knee arthritis who underwent knee arthroscopic synovectomy in our hospital from 2010 to 2017 were enrolled, including 20 patients (16 RA, 4 SpA) with popliteal cyst. Clinical data, RA disease activity score (DAS28), SpA back pain score, etc, were collected to evaluate the efficacy of knee surgery. Results: Erythrocyte sedimentation rate (ESR) [58(17, 79)mm/1h vs. 19(9, 30)mm/1h, P< 0.001],C reactive protein (CRP) [3.72(0.92,8.14) mg/L vs. 0.85(0.10,3.08) mg/L,P<0.001], rheumatoid factor [64.6(20.2,193.3) vs. 20.5(10.0,58.4),P<0.001], DAS28 score(4.67±1.25 vs. 2.81±1.23,P<0.001), knee joint discomfort score [5(4,6) vs. 2(1,3),P<0.001] and the volume of knee joint effusion by ultrasound (P<0.05) in 95 RA patients were significantly decreased compared to those before operation. ESR [27(12,54)mm/1h vs. 20 (16,28) mm/1 h,P<0.001], CRP [3.27(1.06,6.95) mg/L vs. 1.41(0.34,3.03)mg/L,P<0.001],knee discomfort score [2(0,5) vs. 1(0,3),P<0.05], back pain visual analogue score (VAS) [5(4,5) vs. 2(1,3), P<0.001], and the volume of knee joint effusion by ultrasound (P<0.001) in 58 SpA patients were significantly lower than those before the operation.The rate [16.84%(16/95) vs. 6.32%(6/95),P=0.023] and grading (P=0.007) of popliteal cyst in RA were decreased after the operation. No statistically difference was observed in the rate [6.90% (4/58) vs. 5.17%(3/58), P=0.697] of popliteal cyst in patients with SpA, yet with a trend of decrease in 4 patients. Conclusion: This study provide evidence that knee arthroscopic synovectomy has a good effect for refractory knee arthritis, which can reduce disease activity, improve joint symptoms and decrease the grading of popliteal cyst.

[Therapeutic effect of combined use of interferon alpha-1b, interleukin-2 and thalidomide on reversing minimal residual disease in acute myeloid leukemia].

Objective: To explore the effect of combination regimen of interferon alpha-1b, interleukin-2 and thalidomide (ITI regimen) on minimal residual disease (MRD) in patients with acute myeloid leukemia (AML) who were in hematologic remission but MRD-positive. Methods: Eighteen patients (17 from Tumor Hospital of Zhengzhou University and 1 from the First People's Hospital of Pingdingshan City) with AML admitted from July 2016 to June 2018, who were in hematologic remission but MRD-positive were treated with different doses of ITI regimen, and the MRD levels were monitored. Results: Among 18 patients who received a conventional dose of ITI regimen for 1 to 2 months, 7 patients had undetectable MRD, 3 had significant decrease in MRD levels, 3 had elevated MRD level and had hematologic recurrence. Three patients with elevated MRD level received a higher dose of ITI regimen, 2 of them turned to MRD negative and the other 1 patient had decreased MRD level. The total response rate was 72.2%, and the response rate in patients with MRD > 1.0% was 57.1% (4/7) , and that of patients with MRD < 1.0% was 81.8% (9/11) , respectively. Conclusion: The ITI regimen can reduce the MRD level of patient with AML who are in hematologic remission but MRD-positive. The therapeutic effect could be improved by a higher dose administration of ITI regimen, and therapeutic effect may be negatively correlated with MRD level before treatment.

Upregulation of lncRNA LINC00473 promotes radioresistance of HNSCC cells through activating Wnt/ß-catenin signaling pathway.

OBJECTIVE: LncRNA LINC00473 was reported to be upregulated in human cancers. Whereas, the role of LINC00473 in head and neck squamous cell carcinoma (HNSCC) and radiotherapy remains elusive. PATIENTS AND METHODS: Gene array analysis was performed to detect lncRNA LINC00473. Then, the expression of LINC00473 in HNSCC 78 pairs of tissues and cell lines was measured by qRT-PCR assay. To explore the detailed functions of LINC00473 on cell proliferation, MTT and colony formation assays were conducted. We also investigated the influence of LINC00473 expression on radioresistance of HNSCC cells. Western blot assay was used to confirm the relationship between LINC00473 and Wnt/ß-catenin signaling pathway. Furthermore, the effects of x-ray treatment on LINC00473 expression and Wnt/ß-catenin signaling pathway were detected by Western blot assay. RESULTS: LncRNA LINC00473 was upregulated in HNSCC tissues and cell lines. Functional assays showed that LINC00473 acted as oncogene to promote cell proliferation and inhibit apoptosis. In addition, downregulation of LINC00473 enhanced the sensitivity of radiotherapy for HNSCC cells. Furthermore, Western blot assays demonstrated that Wnt/ß-catenin signaling pathway was inhibited by LINC00473 knockdown. Notably, Western blot assay revealed that x-ray treatment suppressed the activity of Wnt/ß-catenin signaling pathway after LINC00473 knockdown. CONCLUSIONS: These data suggested that the upregulation of lncRNA LINC00473 promotes the radioresistance of HNSCC cells through activating Wnt/ß-catenin signaling pathway.

[Risk factors associated with delirium for patients with hip fracture under the orthogeriatric unit mode].

Objective: To explore postoperative delirium (POD)risk factors for geriatric patients who suffered hip fracture under the care of orthogeriatric unit mode. Methods: Patients aged 65 years or older, who were admitted to the orthogeriatric unit in Beijing Jishuitan Hospital from April to October 2017 for hip fracture surgery, were eligible for this prospective cohort study. After univariable analysis, significant risk factors associated with POD were further evaluated with multivariable analysis, to establish independent risk factors associated with POD. Results: A total of 203 patients with an average age of 80(65-96)years were enrolled in the study. The overall incidence of POD was 9.4%(19/203)in which hyperactive and hypoactive type accounting for 84.2%(16/19)and 15.8%(3/19), respectively. Significant difference was found between POD and non-POD groups in patients' age (P=0.003), albumin(P=0.006), TSH(P=0.018), PaCO(2) level(P=0.003), visual analogue scale (VAS )both at rest (P=0.013)and movement(P=0.010) on post-operative day 1.The further Logistic stepwise regression analysis showed that significant differences existed between groups in age(P=0.027), albumin(P=0.003), PaCO(2)(P=0.014)and VAS at rest(P=0.002). Conclusion: The independent risk factors of POD in geriatric patients undergoing hip fracture under the orthogeriatric unit mode include patients' age, pre-operative albumin as well as PaCO(2) level and post-operative VAS at rest.

Estrogen associated gene polymorphisms and their interactions in the progress of Alzheimer's disease.

The extensive neuroprotective effects of estrogen against Alzheimer's disease (AD) have been proven in numerous laboratory studies. However, in clinical studies, the exact role of estrogen in AD is still ambiguous. Some evidences even suggested the high levels of estrogen or estrogen replacement treatment increased the risk of AD. Thus, there must be other factors affecting the neuroprotective effects of estrogen. Multiple enzymes and receptor proteins are involved in the biosynthesis, metabolism and signaling pathways of estrogen, and mediate the beneficial effects of estrogen on AD. Previous studies have suggested some polymorphisms of genes encoding these enzymes and proteins are associated with the risk of AD. In addition to the genes associated with estrogen biosynthesis and metabolism and the genes encoding estrogen receptor proteins, some other genes also modulate the effects of estrogen on AD, or interact with other estrogen-associated genes on the progress of AD. The gene-hormone and gene-gene interactions may be key to unraveling the conflicting results regarding the effect of estrogen on AD. In this paper, we will review and discuss the associations between polymorphisms of these genes and their interactions and the susceptibility to AD. A better understanding of these estrogen-associated genes is significant to explore the pathogenesis of AD.

In vivo DNA-protein interactions at hypersensitive site 3.5 of the human beta-globin locus control region.

Using ligation-mediated polymerase chain reaction and in vivo footprinting methods to study the status of DNA-protein interactions at hypersensitive site 3.5 (HS3.5) of the locus control region in K562 and HEL cells, we found that there was protein occupancy in vivo at HS3.5 in both cell lines and the status of DNA-protein interaction was different between K562 and HEL. These data provide direct evidence that specific nuclear factor-DNA complexes form in vivo at functionally important sequence motifs of the HS3.5 in erythroid cells. This indicates that HS3.5 may play an important role in the regulation of the beta-globin gene cluster. K562 is a human erythroleukemia cell line in which the embryonic epsilon-globin gene is predominantly expressed, while the HEL cell line expresses predominantly the fetal beta-globin genes. Thus, HS3.5 might also be involved in the regulation of developmental stage-specific expression of beta-globin genes. Our results are also consistent with the model that each hypersensitive site acts as a functional unit and HS3.5 may facilitate the formation of the HS3 functional unit.

Both locus control region and proximal regulatory elements direct the developmental regulation of beta-globin gene cluster.

Using ligation-mediated PCR and in vivo footprinting methods to study the status of DNA-protein interaction at hypersensitive site 2 of locus control region and beta(maj) promoter of erythroid cells of fetal liver and adult bone marrow, we found that during different developmental periods, the status of DNA-protein interaction at both hypersensitive site 2 and beta(maj) promoter changed significantly, and indicated that locus control region might function through a looping mechanism to regulate the expression of downstream genes, and that distal regulatory elements (locus control region, hypersensitive sites) as well as proximal regulatory elements (promoter, enhancer) of beta-globin gene cluster participate in the regulation of developmental specificity.

Induction of differentiation and down-regulation of c-myb gene expression in ML-1 human myeloblastic leukemia cells by the clinically effective anti-leukemia agent meisoindigo.

Meisoindigo, a second generation derivative of indirubin, is an effective chemotherapeutic agent with very low toxicity used in the treatment of chronic myeloid leukemia. To determine the nature of this activity, the effect of a nontoxic concentration (0.72 micrograms/mL) of this compound on ML-1 human myeloblastic leukemic cells was examined. At such a concentration, differentiation induction was found to be the most pronounced drug effect. During the 3-day drug incubation period, the viable cell number remained essentially constant, with approximately 48% of the cells demonstrating a mature phenotype with increased acid phosphatase activity and nitroblue tetrazolium dye reduction. As observed with other DNA-specific agents, induction of ML-1 differentiation by meisoindigo was accompanied by the down-regulation of c-myb gene expression. These data suggest that induction of leukemic cell differentiation associated with decreased c-myb expression may be one of the mechanisms of the antitumor action of meisoindigo.

[Vincristine-resistant human KB cell line and mechanism of multidrug resistance].

A multidrug-resistant (mdr) clone of human cancer KB cells was isolated by stepwise selection on exposure to increasing doses of vincristine. The final clone, KBv200, obtained after ethylmethane sulfonate (EMS) mutagenesis showed 175-fold higher resistance to vincristine than did KB cells. The cells were also cross-resistant to taxol, colchicine and adriamycin. Cellular accumulation of vincristine in KBv200 was decreased to less than one-fifth of that in KB. To determine the presence of mdr 1 mRNA in KBv200 and KB, total cellular RNAs from each cell line were analyzed by means of slot blot hybridization. The result showed that the mdr 1 gene had been highly expressed in KBv200. In addition, verapamil, a calcium channel blockers, was shown to increase VCR accumulation in KBv200 and reverse the vincristine resistance. All these results demonstrate that the mechanism of KBv200 cell resistance to multiple drugs resulted from increased expression of mdr 1 gene and brought about over production of P-glycoprotein and increased the efflux of drugs.

[The cytotoxicity and action mechanism of ranunculin in vitro].

This paper describes the cytotoxicity of ranunculin (RAN) and its mechanism of action. The IC50 of RAN against the KB and Bel7402 cells in colony test were found to be 0.21 and 0.35 mumol/L respectively. RAN inhibited the incorporation of 3H-labeled precursors into DNA and RNA of L1210 cells. RAN (15 mumol/L) markedly decreased DNA synthesis catalyzed by DNA polymerase I and promoted the generation of superoxide anions in DMSO/KO2 system. In the meantime, SOD and CAT were shown to partly revoke the inhibitory effects of RAN upon the incorporation of 3H-TdR into DNA. No direct reaction between RAN and DNA template was observed and no effect of RAN on DNA TOPO II or RNA polymerase was found. Our results suggest that the cytotoxicity of RAN in vitro may be due to inhibition of DNA polymerase and increase of oxygen free radicals.

[Antimutagenic activity and metabolic transformation of ranunculin by rat liver microsomes].

Ranunculin (RAN) was shown to be an antimutagenic agent selectively against mitomycin C (MMC) or methyl methane sulfanate (MMS) treated Salmonella typhimminum TA100/TA102. It decreased the formation of micronucleus of MMC induced polychromatic erythrocytes (PEC) from 46 +/- 9.2% to 20 +/- 6% in mice. The inhibition of RAN on the incorporation of 3H-TdR into DNA disappeared after incubation with rat liver microsomes and cytoplasm since RAN was found to be metabolized by rat liver microsomes in vitro, resulting in a new absorbance peak at 258 nm, determined by RP-HPLC. The data show that RAN may have both antimutagen and antitumor activity, but the latter action may disappear by metabolic transformation.

Pharmacological studies of meisoindigo: absorption and mechanism of action.

Meisoindigo, an indirubin derivative, is a new type of cancer chemotherapeutic agent. It exhibited higher activity against rodent tumors than indirubin itself. Experiments have shown the improved absorption of meisoindigo, compared to indirubin to be one of the major reasons for the enhancement of antitumor activity. Studies on the mechanism of meisoindigo action indicate that it strongly inhibits DNA biosynthesis in tumor cells. Strong inhibition of the drug on assembly of microtubule protein was also obtained. By means of FCM technique the effects of meisoindigo on mouse leukemia L1210 cell cycle were examined. Experimental results showed that under the action of meisoindigo the S phase cells accumulated and the traverse of the cells in G2 + M phase to G1 phase may also be blocked to some extent.

Determination of DNA topoisomerase II activity from L1210 cells--a target for screening antitumor agents.

DNA topoisomerase II was isolated from mouse leukemia L1210 cells and the activity was determined by using P4 phage knotted DNA and pBR 322 DNA as the substrates. Based on these results, a method for screening antitumor agents by using DNA topoisomerase II as a target was established. The experiments showed that DNA topoisomerase II catalyzed pBR 322 DNA breaking and relaxing which were reversible and dependent on ATP. The activity was increased 2-4 times in the presence of ATP 1 mmol.L-1. In contrast with type II enzyme, the activity of DNA topoisomerase I was completely inhibited in the presence of ATP 1 mmol.L-1 and had full activity in the absence of ATP. Type II enzyme also showed the unknotting activity by using p4 phage knotted DNA as a substrate. DNA cleavage and relaxing reaction induced by type II enzyme increased 5-fold in the presence of Doxorubicin (Dox) 1 microgram.ml-1 or daunorubicin (Dau). Etoposide (Eto) and aclarubicin B (Acl B) also stimulated the reaction at 100 micrograms.ml-1. The cleavage reaction resulted from topoisomerase II was inhibited by other agents, such as frankincense extracts, terpenic compounds (BC series).

[Depletion of O6-alkylguanine alkyltransferase and chromosome damage induced by cisplatin, ning xin platin and carboplatin].

O6-Alkylguanine-DNA-alkyltransferase (O6-AGT) is a very important DNA repair protein known to carry out the transfer of alkyl groups from the O6 position of guanine in alkylated DNA to a cysteine acceptor site contained within its own protein sequence. In this work, the activity of O6-AGT in different cell lines and the relationship between the depletion of the enzyme and the frequency of micronuclei induced by cisplatin (DDP), Ning Xin platin (camphoramine chloroacetic platinum, CCP) or carboplatin (JM-8) in KB and CHL cells were studied. Experiments indicate that KB cells showed higher O6-AGT activity (greater than 400 dpm/300 micrograms protein extracts) which belonged to Mer+ cells, but CHL, HL-60 and L1210 cells showed very low O6-AGT activity (less than 50 dpm/300 micrograms protein extracts) which can be considered to be Mer- cells. Cytotoxicity studies indicated that no mer- selection was observed in these platinum complexes for KB, HL-60, CHL and L1210 cells. However, a good relationship between the depletion of O6-AGT and the frequency of micronuclei induced by the platinum complexes was obtained. CCP caused the highest depletion of the enzyme and exhibited highest potency in damaging chromosome.

Establishment and biological properties of mouse granulocytic leukemic cell line-L833.

A granulocytic leukemic cell line, called L833 has been established through culturing in vitro from mouse bone marrow of transplantable granulocytic leukemia. More than 280 passages have been performed in 3 years. Cell growth has been rapid and stable. Incidence of tumour formation was 100% with various routes of inoculation. The nature of granulocytic leukemia was confirmed by examination of cytology, cytochemistry, pathology and ultrastructural changes. Analysis proved chromosomes to be hypodiploid with model number of 39, loss of chromosomes, and presence of a marker. Besides the chromosomal change as mentioned above, both L883-A and L833-B derived from colonies formed from the L833 cells cultured in semi-solid agar medium, have their own marker. The cell line was sensitive to various types of antitumour agents in varying degrees. It was also sensitive to ionizing radiation. D0 values of L833 and L883-A cells were 98.8 and 104.9 rad, respectively. The results were similar to that of L801. Establishment of this cell line is of important significance for studying leukemia and screening anti-tumour agents, as well as provides a useful direct, economic and precisely quantitative tool for other relative studies.

Establishment of a model of transplantable myelocytic leukemia (L801) in LACA mice.

A transplantable myelocytic leukemia model of LACA mice, designated by the name of L801, was established by intravenous injection of spleen cell suspension from mice with radiation-induced myelocytic leukemia into mice of the same strain. Until now, for more than three years, the L801 has maintained stable and rapid growth and has been reproduced for over 130 serial passages. The incidence of leukemia in inoculated animals was approximately 100% and mean survival time was 10.9 +/- 2.1 days. The L801 is of myelocytic type which has been determined by cytological, cytochemical, pathological and ultrastructural observations. Its karyotype was hypodiploid, characterized by modal number of 39, loss of Y chromosome and an abnormal huge marker chromosome. The cell cycle duration of the L801 was 16 h. C-type viral particles were observed under the electron-microscope. The L801 was sensitive, to varying extents, to various anti-tumor agents. We presume that the L801 is a useful tool in studies on mechanism of leukemogenesis, anti-tumor agent screening and treatment of experimental tumors.