Inducement of ER Stress by PAD Inhibitor BB-Cl-Amidine to Effectively Kill AML Cells.

OBJECTIVE: Acute myeloid leukemia (AML) is a highly heterogeneous and recurrent hematological malignancy. Despite the emergence of novel chemotherapy drugs, AML patients' complete remission (CR) remains unsatisfactory. Consequently, it is imperative to discover new therapeutic targets or medications to treat AML. Such epigenetic changes like DNA methylation and histone modification play vital roles in AML. Peptidylarginine deminase (PAD) is a protein family of histone demethylases, among which the PAD2 and PAD4 expression have been demonstrated to be elevated in AML patients, thus suggesting a potential role of PADs in the development or maintenance of AML and the potential for the identification of novel therapeutic targets. METHODS: AML cells were treated in vitro with the pan-PAD inhibitor BB-Cl-Amidine (BB-Cl-A). The AML cell lines were effectively induced into apoptosis by BB-Cl-A. However, the PAD4-specific inhibitor GSK484 did not. RESULTS: PAD2 played a significant role in AML. Furthermore, we found that BB-Cl-A could activate the endoplasmic reticulum (ER) stress response, as evidenced by an increase in phosphorylated PERK (p-PERK) and eIF2α (p-eIF2α). As a result of the ER stress activation, the BB-Cl-A effectively induced apoptosis in the AML cells. CONCLUSION: Our findings indicated that PAD2 plays a role in ER homeostasis maintenance and apoptosis prevention. Therefore, targeting PAD2 with BB-Cl-A could represent a novel therapeutic strategy for treating AML.

Determination of Biogenic Amines in Different Parts of Lycium barbarum L. by HPLC with Precolumn Dansylation.

The aim of this work was to characterize biogenic amines (BAs) in different parts of Lycium barbarum L. using HPLC with dansyl chloride derivatization, and jointly, to provide referential data for further exploration and utilization of Lycium barbarum L. The linear correlation coefficients for all BAs were above 0.9989. The limits of detection and quantification were 0.015-0.075 and 0.05-0.25 µg/mL, respectively. The relative standard deviations for the intra-day and inter-day precision were 0.66-2.69% and 0.91-4.38%. The described method has good repeatability and intermediate precision for the quantitative determination of BAs in different parts of Lycium barbarum L. Satisfactory recovery for all amines was obtained (79.3-110.3%). The result showed that there were four kinds of BAs. The highest putrescine content (20.9 ± 3.2 mg/kg) was found in the flower. The highest histamine content (102.7 ± 5.8 mg/kg) was detected in the bark, and the highest spermidine (13.3 ± 1.6 mg/kg) and spermine (23.7 ± 2.0 mg/kg) contents were detected in the young leaves. The high histamine (HIS) content in the bark may be one of the reasons why all of the parts of Lycium barbarum L., except the bark, are used for medicine or food in China. Meanwhile, the issue of the high concentration of HIS should be considered when exploiting or utilizing the bark of Lycium barbarum L.

Incidence and risk factors of tuberculosis among the elderly population in China: a prospective cohort study.

BACKGROUND: China is facing challenges of the shifting presentation of tuberculosis (TB) from younger to elderly due to an ageing population, longer life expectancy and reactivation disease. However, the burden of elderly TB and influence factors are not yet clear. To fill the gap, we generated a cohort study to measure the magnitude of TB incidence and associated factors among the elderly population aged 65 years and above in China. METHODS: In this cohort established in 2013 through a prevalence survey conducted in selected sites, a total of 34 076 elderlies without TB were enrolled into two-year follow-up. We used both active and passive case findings to find out all TB patients among them. The person-year (PY) incidence rates for both bacteriologically positive TB and active TB were calculated. Cox proportional regression model was performed to test effect of risk factors, and the population attributable fraction (PAF) of each risk factor contributing to incident TB among elderlies was calculated. RESULTS: Over the two-year follow-up period, a total of 215 incident active TB were identified, 62 of which were bacteriologically positive. The incidence rates for active TB and bacteriologically positive TB were 481.8 per 100 000 PY (95% CI: 417.4-546.2 per 100 000 PY) and 138.9 per 100 000 PY (95% CI: 104.4-173.5 per 100 000 PY), respectively. Incident cases detected by active case finding were significantly higher (P < 0.001). Male, non-Han nationality, previously treated TB, ex/current smoker and body mass index (BMI) < 18.5 presented as independent predictors for developing TB disease. For developing bacteriologically positive TB, the biggest contribution was from self-reported ex or current smoker (18.06%). And, for developing active TB, the biggest contribution was from non-Han nationality (35.40%), followed by male (26.80%) and age at 75 years and above (10.85%). CONCLUSIONS: Ageing population in China had a high TB incidence rate and risk to develop TB disease, implying that National TB Program (NTP) needs to prioritize for elderly. Active case finding should be applied capture more active TB cases among this particular population, especially for male, non-Han nationality, and those with identified risk factors.

Resveratrol: Review on its discovery, anti-leukemia effects and pharmacokinetics.

Resveratrol, found in variety of plants, is a natural stilbene structure polyphenol. It has various pharmacological effects, such as antioxidation, anti-aging, anti-inflammation, anti-cancer, antiobesity, anti-diabetes, cardioprotection, neuroprotection. Recently, anti-leukemia activities of resveratrol has been studied extensively via its effects on a variety of biological processes involving cell proliferation, apoptosis, autophagy. Current treatments of leukemia mainly rely on intensive chemotherapy or hematopoietic stem cell transplantation, however, these treatments are still with poor survival and high treatment-related mortality. Therefore, it is extremely needed to find relatively non-toxic medicines with minimal side effects but sufficient therapeutic efficacy. Resveratrol is one such potential candidate owing to its reported anti-leukemia effect. In this review, we summarized resveratrol's discovery, sources and isolation methods, administration methods, effects in different types of leukemia, pharmacokinetics and toxicities, aiming to exploit resveratrol as a potential drug candidate for anti-leukemia.

Arginine vasopressin differentially modulates GABAergic synaptic transmission onto temperature-sensitive and temperature-insensitive neurons in the rat preoptic area.

The preoptic area (POA) of the hypothalamus, containing temperature-sensitive and temperature-insensitive neurons, plays a key role in specific thermoregulatory responses. Although arginine vasopressin (AVP) has been shown to induce hypothermia by increasing the firing activities of warm-sensitive neurons and decreasing those of cold-sensitive and temperature-insensitive neurons, the effects of AVP on POA GABAergic transmission remain unknown. Herein, inhibitory postsynaptic currents (IPSCs) of temperature-sensitive and temperature-insensitive neurons in POA slices were recorded using whole-cell patch clamp. By monitoring changes in GABAergic transmission during AVP treatment, we showed that AVP decreased the amplitudes and frequencies of spontaneous IPSCs in mostly warm-sensitive neurons and in some temperature-insensitive neurons but increased these parameters in other temperature-insensitive neurons. The IPSC amplitude was reduced for only cold-sensitive neurons. RT-PCR and Western blot analyses further confirmed the POA expression of V1a receptors and GABAA receptors, including the subunits α1, α2, α3, ß2, ß3 and γ2. The effects of AVP on IPSCs in temperature-sensitive and temperature-insensitive neurons were dependent on G proteins and intracellular Ca2+ . AVP-mediated modulation was associated with changes in the kinetic parameters (decay time, 10-90% rise time, half-width). Together, these results suggest that AVP, acting via V1a receptors but not V1b receptors, differentially modulates GABAergic synaptic transmission and fine-tunes the firing activities of temperature-sensitive and temperature-insensitive neurons in the rat POA.

Improving tuberculosis case detection in underdeveloped multi-ethnic regions with high disease burden: a case study of integrated control program in China.

BACKGROUND: In the underdeveloped multi-ethnic regions of China, high tuberculosis (TB) burden and regional inequity in access to healthcare service increase the challenge of achieving the End TB goals. Among all the provinces, the highest TB burden is reported in Xinjiang, where ethnic minorities and older people have suffered most. However, current case-finding strategy is inadequate given the complex social determinants and suboptimal case detection rates. Thus, we developed an integrated TB control program to improve case detection and conducted a pilot in Xinjiang from 2014 to 2015. In this case study, we summarized the activities and key findings. We also shared the experiences and challenges of implementing interventions and provided recommendations to inform the TB control program in the future. CASE PRESENTATION: The pilot interventions were implemented in one selected town in Yining based on local TB control programs. By applying tailor-made educational materials, outreach TB educational activities were conducted in diverse ways. In 22 Masjids, the trained imams promoted TB education to the Muslims, covering 20,440 person-times in 88 delivered preaching sessions. In seven schools, 1944 students were educated by the teachers and contributed to educating 6929 family members. In the village communities, 13,073 residents participated in household education and screening. Among them, 12,292 people aged under 65 years were investigated for suspicious pulmonary TB symptoms, where six TB patients were diagnosed out of 89 TB suspects; 781 older people were mobilized for screening directly by chest X-ray, where 10 patients were diagnosed out of 692 participants. Supportive healthcare system, multi-sectoral cooperation and multi-channel financing mechanism were the successful experiences of implementation. The interventions were proved to be more effective than the previous performance: the number of TB suspects consulting doctors and patients detected increased by 50% and 26%, respectively. The potential challenges, implications and recommendations should been taken into account for further program improvement. CONCLUSIONS: In underdeveloped multi-ethnic regions with high TB burden, improving case detection is necessary and the interventions can be feasible and effective within a supportive system. More intensive educational and training approaches, a high index of TB suspicion and prioritization of older people in screening are recommended. To sustain and scale up the program, the impacts, cost-effectiveness, feasibility and acceptability of interventions warrant further research and evaluation in each specific context.

Photo-protective mechanisms in reed canary grass to alleviate photo-inhibition of PSII on the Qinghai-Tibet Plateau.

Due to its characteristic of high biomass yield potential, there is considerable interest in cultivating Phalaris arundinacea L. cv. 'chuancaoyin No.3' (reed canary grass) on the Qinghai-Tibet Plateau where there is an abundance of alpine steppe meadow and a potential large market for animal husbandry. In this study, we 1) investigate whether reed canary grass exhibits superior productive capacity to Elymus nutans 'Aba' (E. nutans), ordinary common pasture, during the long warm days of summer at high-altitude; and 2) compare the cold tolerance between reed canary grass and E. nutans, including photosynthesis, photo-inhibition, and photo-protection. The results suggest that reed canary grass exhibits higher photosynthetic capacity compared to E. nutans at latitudes of the cool temperate zone. Meanwhile, cold-induced photo-inhibition and photo-damage at high altitudes in reed canary grass were due to both stomatal and non-stomatal limitation, and the enhancement in photo-respiration, thermal dissipation, and Mehler reaction are important processes to minimize the negative effects of high elevation and a cold environment.

Resveratrol attenuates skeletal muscle atrophy induced by chronic kidney disease via MuRF1 signaling pathway.

Skeletal muscle atrophy is an important clinical characteristic of chronic kidney disease (CKD); however, at present, the therapeutic approaches to muscle atrophy induced by CKD are still at an early stage of development. Resveratrol is used to attenuate muscle atrophy in other experimental models, but the effects on a CKD model are largely unknown. Here, we showed that resveratrol prevented an increase in MuRF1 expression and attenuated muscle atrophy in vivo model of CKD. We also found that phosphorylation of NF-κB was inhibited at the same time. Dexamethasone-induced MuRF1 upregulation was significantly attenuated in C2C12 myotubes by resveratrol in vitro, but this effect on C2C12 myotubes was abrogated by a knockdown of NF-κB, suggesting that the beneficial effect of resveratrol was NF-κB dependent. Our findings provide novel information about the ability of resveratrol to prevent or treat muscle atrophy induced by CKD.

Effects of mineralocorticoid receptor antagonists on the progression of diabetic nephropathy.

AIMS/INTRODUCTION: We aimed to evaluate the potential benefits and adverse effects of adding a mineralocorticoid receptor antagonist (MRA) to angiotensin-converting enzyme inhibitors (ACEI) and/or angiotensin receptor blockers (ARB), as standard treatment in patients with diabetic nephropathy. MATERIALS AND METHODS: We scanned the Embase, PubMed and Cochrane Central Register of Controlled Trials databases for human clinical trials published in English until June 2016, evaluating renal outcomes in patients with diabetic nephropathy. RESULTS: A total of 18 randomized controlled trials involving 1,786 patients were included. Compared with ACEI/ARB alone, co-administration of MRA and ACEI/ARB significantly reduced urinary albumin excretion and the urinary albumin-creatinine ratio (mean difference -69.38, 95% confidence intervals -103.53 to -35.22, P < 0.0001; mean difference -215.74, 95% confidence intervals -409.22 to -22.26, P = 0.03, respectively). A decrease of blood pressure was also found in the co-administration of MRA and ACEI/ARB groups. However, we did not observe any improvement in the glomerular filtration rate. There was a significant increase in the risk of hyperkalemia on the addition of MRA to ACEI/ARB treatment (relative risk 3.74, 95% confidence intervals 2.30-6.09, P < 0.00001). CONCLUSIONS: These findings suggest that co-administration of MRA and ACEI/ARB has beneficial effects on renal outcomes with increasing the incidence of hyperkalemia.

Medicago truncatula genotypes Jemalong A17 and R108 show contrasting variations under drought stress.

Drought is one of the most significant abiotic stresses that restrict crop productivity. Medicago truncatula is a model legume species with a wide genetic diversity. We compared the differential physiological and molecular changes of two genotypes of M. truncatula (Jemalong A17 and R108) in response to progressive drought stress and rewatering. The MtNCED and MtZEP activation and higher abscisic acid (ABA) content was observed in Jemalong A17 plants under normal conditions. Additionally, a greater increase in ABA content and expression of MtNCED and MtZEP in Jemalong A17 plants than that of R108 plants were observed under drought conditions. A more ABA-sensitive stomatal closure and a slower water loss was found in excised leaves of Jemalong A17 plants. Meanwhile, Jemalong A17 plants alleviated leaf wilting and maintained higher relative water content under drought conditions. Exposed to drought stress, Jemalong A17 plants exhibited milder oxidative damage which has less H2O2 and MDA accumulation, lower electrolyte leakage and higher chlorophyll content and PSII activity. Furthermore, Jemalong A17 plants enhanced expression of stress-upregulated genes under drought conditions. These results suggest that genotypes Jemalong A17 and R108 differed in their response and adaptation to drought stress. Given the relationship between ABA and these physiological responses, the MtNCED and MtZEP activation under normal conditions may play an important role in regulation of greater tolerance of Jemalong A17 plants to drought stress. The activation of MtNCED and MtZEP may lead to the increase of ABA content which may activate expression of drought-stress-regulated genes and cause a series of physiological resistant responses.

Synthesis of new 4-anilinoquinazoline analogues and evaluation of their EGFR inhibitor activity.

Thirteen of 4-anilinoquinazoline derivatives with imine groups at position 6 of quinazoline ring were synthesized and their antitumor activities were evaluated by MTT assay and Western blotting analysis. Among these compounds, 13a-131 were reported first time. The MTT assay was carried out on three human cancer cell lines (A549, HepG2 and SMMC7721) with EGFR highly expressed. Among the tested compounds, 13i and 13j exhibited notable inhibition potency and their IC50 values on three cell lines were equivalent to or less than those of gefitinib. Compound 14, without imine group substituted, displayed excellent inhibitor potency only on A549 cell line. Compounds 14 and 13j were chosen to perform Western blotting analysis on A549. The results showed that both of the compounds could inhibit the expression level of phosphorylated EGFR remarkably. It was concluded that the inhibitor potency of compound 14 was almost equivalent to that of gefitinib and the inhibitor potency of 13j was better than that of gefitinib.

Benign giant-cell tumor of the common bile duct: a case report.

Primary giant-cell tumors rarely arise in the common bile duct. We herein report a case of primary giant-cell tumor of the common bile duct. The patient was an 81-year-old male who was diagnosed with a well-defined 1.2-cm mass projecting into the lumen of the middle common bile duct. Excision of the gallbladder and extrahepatic bile duct and a Roux-en-Y cholangiojejunostomy were performed. Histologically, the tumor had no association with carcinomas of epithelial origin and was similar to giant-cell tumors of the bone. The tumor consisted of a mixture of mononuclear and multinucleated osteoclast-like giant cells. The mononuclear cells showed no atypical features, and their nuclei were similar to those of the multinucleated giant cells. CD68 was expressed on the mononuclear and multinucleated osteoclast-like giant cells, whereas CD163 immunoreactivity was restricted to the mononuclear cells. Six months after the operation, the patient was still alive and had no recurrence. The interest of this case lies in the rarity of this entity, the difficulty of preoperative diagnosis, and this tumor's possible confusion with other malignant tumors.

[The long term (9-year) survival of multidrug-resistant tuberculosis patients compared to non-multidrug-resistant tuberculosis patients in Henan province].

OBJECTIVE: To investigate the long term survival of MDR-TB patients compared to non-MDR-TB in Henan province in 2010. METHODS: Participants were randomly selected in 2010 from a dataset generated by an anti-TB drug resistance surveillance survey conducted by the Tuberculosis Control Institute, Henan Centre for Disease Control and Prevention in 2001, supported by the World Health Organization. Information on patient's demographic profile and medical records was extracted by trained doctors and nurses at local anti-TB dispensaries. Interviews were carried out using questionnaires to collect information on the socioeconomic features and survival status. Bivariate and multivariate with logistic regression were performed for data analysis. RESULTS: The long term outcome of MDR-TB patients was much poorer when compared to non-MDR-TB patients. The case fatality was much higher among MDR-TB than non-MDR-TB patients (22.1% vs. 6.7%). The risk factors associated with the poorer outcome would include drug resistance status, disease relapse, hospitalization for treatment and long treatment period. Compared to non-MDR-TB, the survival time for MDR-TB was much shorter after having had the disease (6.7 years vs. 8.0 years). CONCLUSION: MDR-TB patients had poor long term outcomes. As most of the cured TB patients were under productive age in the society, the high case fatality rate of MDR-TB would impose big burden on the related family and communities. Findings from this study suggested that the TB control programs should involve more efforts be paid on MDR-TB control, in order to reduce the burden of the disease.

Activation of satellite glial cells in lumbar dorsal root ganglia contributes to neuropathic pain after spinal nerve ligation.

The role of satellite glial cells (SGCs) of sensory ganglia in chronic pain begins to receive interest. The present study aims to investigate the contribution of SGC activation to the development of neuropathic pain. A neuropathic pain model was established by lumbar 5 spinal nerve ligation (SNL), and glial fibrillary acidic protein (GFAP) was used as a marker of SGC activation. It was found that SGCs were activated in the ipsilateral dorsal root ganglia (DRG) increased significantly as early as 4h following SNL, gradually increased to a peak level at day 7, and then stayed at a high level to the end of the experiment at day 56. SGC activation in the SNL group was significantly higher than that in the sham group at days 1, 3 and 7 after operation. Immunofluorescent double labeling showed that the activated SGCs encircled large, medium-sized and small neurons. The SGCs surrounded the small and medium-sized neurons were preferentially activated in the early phase, but shifted to large diameter neurons as time went on. Continuous infusion of fluorocitrate, a glial metabolism inhibitor, to the affected DRG via mini-osmotic pump for 7d significantly alleviated mechanical allodynia at day 7. These results suggest that SGCs in the DRG were activated after SNL. SGC activation contributed to the early maintenance of neuropathic pain.

Role of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor subunit GluR1 in spinal dorsal horn in inflammatory nociception and neuropathic nociception in rat.

The present study aims to investigate changes of spinal cord AMPA receptor GluR1 and its phosphorylation in inflammatory and neuropathic pain. Complete Freund's adjuvant (CFA) injection into the hind paw produced inflammatory thermal hyperalgesia that was assessed by decreased response latency to radiant heat; spinal nerve ligation (SNL) was used to induce mechanical allodynia that was evaluated with von Frey hairs. By method of Western blot, expression of GluR1 (the main subunit of the AMPA receptor) and its phosphorylated forms at serine 845 (pGluR1-Ser845) and at serine 831 (pGluR1-Ser831) in the spinal dorsal horn was observed. It was found that the expression of pGluR1-Ser845 and pGluR1-Ser831 increased significantly at 1 h after CFA injection, reached peak at 4 h and returned to the normal control level at 24 h, while no significant change was detected in GluR1 itself. In contrast, neither GluR1 nor pGluR1 showed any significant change in rats following SNL. These results suggest that phosphorylated GluR1 (pGluR1-Ser845 and pGluR1-Ser831) might play a role in the induction of inflammatory but not neuropathic pain.