[The Effect of SIRT5 Deletion on Recovery of Hematopoietic Stem Cells after Injury in Mouse].

OBJECTIVE: To investigate the effect of deacylase Sirtuin 5 in the recovery of hematopoietic stem cells (HSCs) after treated by 5-FU in mouse. METHODS: Flow cytometry was used to analyze the effect of SIRT5 deletion on the proportion of hematopoietic stem/progenitor cells (HSPCs) in bone marrow (BM), the proportion of T cells, B cells and myeloid cells (TBM) in peripheral blood (PB) and spleen, and the development of T cells in thymus. Mouse were treated with 5-FU to study the effect of SIRT5 deletion on the cell cycle, apoptosis and the proportion of HSPCs in BM. The effect of SIRT5 deletion on the proliferation of HSCs was analyzed by flow sorting in vitro. RESULTS: SIRT5 deletion did not affect the development of T cells in thymus and the proportion of TBM cells in PB and spleen compared with wild type mice. SIRT5 deletion increased proportion of HSPCs in BM. After 5-FU treatment, the proportion of HSCs in SIRT5 deletion mice was significant decreased (P < 0.05), the HSPC in SIRT5 deletion mice was activated from G0 to G1 phase (P < 0.05), and the proportion of early apoptosis increased (P < 0.05). By monoclonal culture in vitro, the ability of HSCs to form clones in SIRT5 deletion mice was decreased significantly (P < 0.05). CONCLUSION: SIRT5 deletion lead to a decreased the ability of HSCs to clone in vitro. SIRT5 deletion is not conducive to the recovery of HSPCs injury in mice under hematopoietic stress.

Small non-coding RNAome changes during human chondrocyte senescence as potential epigenetic targets in age-related osteoarthritis.

Chondrocyte senescence is a decisive component of age-related osteoarthritis, however, the function of small noncoding RNAs (sncRNAs) in chondrocyte senescence remains underexplored. Human hip joint cartilage chondrocytes were cultivated up to passage 4 to induce senescence. RNA samples were extracted and then analyzed using small RNA sequencing and qPCR. ß-galactosidase staining was used to detect the effect of sncRNA on chondrocyte aging. Results of small RNA sequencing showed that 279 miRNAs, 136 snoRNAs, 30 snRNAs, 102 piRNAs, and 5 rasiRNAs were differentially expressed in senescent chondrocytes. The differential expression of 150 sncRNAs was further validated by qPCR. Transfection of sncRNAs and ß-galactosidase staining were also performed to further revealed that hsa-miR-135b-5p, SNORA80B-201, and RNU5E-1-201 have the function to restrain chondrocyte senescence, while has-piR-019102 has the function to promote chondrocyte senescence. Our data suggest that sncRNAs have therapeutic potential as novel epigenetic targets in age-related osteoarthritis.

Reducing host aldose reductase activity promotes neuronal differentiation of transplanted neural stem cells at spinal cord injury sites and facilitates locomotion recovery.

Neural stem cell (NSC) transplantation is a promising strategy for replacing lost neurons following spinal cord injury. However, the survival and differentiation of transplanted NSCs is limited, possibly owing to the neurotoxic inflammatory microenvironment. Because of the important role of glucose metabolism in M1/M2 polarization of microglia/macrophages, we hypothesized that altering the phenotype of microglia/macrophages by regulating the activity of aldose reductase (AR), a key enzyme in the polyol pathway of glucose metabolism, would provide a more beneficial microenvironment for NSC survival and differentiation. Here, we reveal that inhibition of host AR promoted the polarization of microglia/macrophages toward the M2 phenotype in lesioned spinal cord injuries. M2 macrophages promoted the differentiation of NSCs into neurons in vitro. Transplantation of NSCs into injured spinal cords either deficient in AR or treated with the AR inhibitor sorbinil promoted the survival and neuronal differentiation of NSCs at the injured spinal cord site and contributed to locomotor functional recovery. Our findings suggest that inhibition of host AR activity is beneficial in enhancing the survival and neuronal differentiation of transplanted NSCs and shows potential as a treatment of spinal cord injury.

Target inhibition of caspase-8 alleviates brain damage after subarachnoid hemorrhage.

Caspase-8 plays an important role in the mediation of inflammation and the effect of its role in subarachnoid hemorrhage remains elusive. The nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome has been postulated to mediate inflammation during SAH. The aim of the present study was to investigate the effects of caspase-8 inhibition on SAH injury and further elucidate the molecular mechanisms. In this study, a subarachnoid hemorrhage model was established by endovascular perforation process in adult male Sprague-Dawley rats. Z-IETD-FMK (0.5, 1, 2 mg/kg; an inhibitor of caspase-8) was delivered via intravenous (tail vein) injection immediately after subarachnoid hemorrhage. After 12 hours of subarachnoid hemorrhage, western blot assay showed that the expression of cleaved caspase-8 was significantly increased at 12 hours, peaked at 24 hours, and then decreased at 72 hours after subarachnoid hemorrhage. Immunofluorescence staining demonstrated that caspase-8 was expressed in microglia after subarachnoid hemorrhage. Z-IETD-FMK significantly improved neurological deficits and reduced brain water content 24 hours after subarachnoid hemorrhage. The Morris water maze and rotarod test confirmed that Z-IETD-FMK significantly improved spatial learning and memory abilities and motor coordination at 21-27 days after subarachnoid hemorrhage. Furthermore, inhibition of caspase-8 activation reduced the expression of pyrin domain-containing 3, caspase-1, and interleukin-1ß after subarachnoid hemorrhage. In conclusion, our findings suggest that caspase-8 inhibition alleviates subarachnoid hemorrhage-induced brain injuries by suppressing inflammation. The study was approved by the Institutional Animal Ethics Committee of the First Affiliated Hospital, School of Medicine, Zhejiang University, China (approval No. 2016-193) on February 25, 2016.

Exciplex Cohosts Employing Nonconjugated Linked Dicarbazole Donors for Highly Efficient Thermally Activated Delayed Fluorescence-Based Organic Light-Emitting Diodes.

Two new nonconjugated linked dicarbazole materials, dCzPSi and dCzPSO2, with high triplet energy were synthesized and characterized. dCzPSi and dCzPSO2 were adopted as unipolar host materials for the green thermally activated delayed fluorescence (TADF) emitter (4CzIPN) to achieve high-efficiency organic light-emitting diodes (OLEDs). The electron-transporting acceptor, PO-T2T, was introduced to mix with dCzPSi and dCzPSO2 to give two new exciplex-forming systems that can improve the exciton formation propensity in the emitting layer. The relevant properties of these new exciplexes were characterized, and they were suggested as promising cohosts for the green TADF emitter, 4CzIPN. The characteristics of the devices employing single hosts (dCzPSi and dCzPSO2) and exciplex-forming cohosts (dCzPSi:PO-T2T and dCzPSO2:PO-T2T) were explored. The obtained results indicate that the Si-bridged dicarbazole compound dCzPSi outperforms its counterpart dCzPSO2 in which two carbazole groups are linked by an SO2 group. The device employed with the dCzPSi:PO-T2T cohost with 10 wt % 4CzIPN achieved a low Von of 2.2 V and maximum efficiencies of 21.1% (external quantum efficiency), 56.4 cd A-1 (current efficiency), 59.1 lm W-1 (power efficiency), as compared to those (18.7%, 56.6 cd A-1, and 68.5 lm W-1) of the dCzPSO2:PO-T2T-hosted device. This work verifies the advantages of using a cohost that can form an exciplex for boosting the device efficiency with reduced efficiency roll-off of TADF-based OLEDs.

Rapamycin inhibition of eosinophil differentiation attenuates allergic airway inflammation in mice.

BACKGROUND AND OBJECTIVE: The mammalian target of rapamycin (mTOR) signalling pathway regulates immune responses, and promotes cell growth and differentiation. Inhibition of mTOR with rapamycin modulates allergic asthma, while the underlying molecular mechanisms remain elusive. Here, we demonstrate that rapamycin, effectively inhibits eosinophil differentiation, contributing to its overall protective role in allergic airway inflammation. METHODS: Rapamycin was administered in a mouse model of ovalbumin-induced allergic airway inflammation, and the eosinophil differentiation was analysed in vivo and in vitro. RESULTS: Rapamycin significantly attenuated allergic airway inflammation and markedly decreased the amount of eosinophils in local airways, peripheral blood and bone marrow, independently of levels of interleukin-5 (IL-5). In vitro colony forming unit assay and liquid culture demonstrated that rapamycin directly inhibited IL-5-induced eosinophil differentiation. In addition, rapamycin reduced the production of IL-6 and IL-13 by eosinophils. Rapamycin was also capable of reducing the eosinophil levels in IL-5 transgenic NJ.1638 mice, again regardless of the constitutive high levels of IL-5. Interestingly, rapamycin inhibition of eosinophil differentiation in turn resulted in an accumulation of eosinophil lineage-committed progenitors in bone marrow. CONCLUSIONS: Altogether these results clearly demonstrate a direct inhibitory role of rapamycin in eosinophil differentiation and function, and reemphasize the importance of rapamycin and possibly, mTOR, in allergic airway disease.

Effect of intravenous general anaesthesia with epidural block on the expression of pre-endogenous opioid peptide genes.

OBJECTIVE: To measure the plasma concentrations of three endogenous opioid peptides and the levels of preproenkephalin (PPE) and preprodynorphin (PPD) mRNA in peripheral blood lymphocytes of patients during scheduled surgery performed under intravenous general anaesthesia combined with an epidural block. METHODS: Patients were anaesthetized and arterial blood was collected at 0 (baseline), 20, 40, 60, and 80 min during surgery. The plasma concentrations of ß-endorphin, leucine-enkephalin and dynorphin A were measured using radioimmunoassay. Reverse transcription-polymerase chain reaction was used to measure the levels of PPD and PPE mRNA in peripheral blood lymphocytes collected during surgery. RESULTS: Fifteen patients participated in this prospective study. The plasma concentrations of ß-endorphin were significantly lower at all time-points compared with the baseline value. The plasma concentrations of leucine-enkephalin and dynorphin A were significantly lower at 40, 60, and 80 min compared with baseline. The PPD/ß-actin ratio was significantly lower at 80 min compared with baseline, while the PPE/ß-actin ratio showed no significant change. CONCLUSION: The level of mRNA from two pre-endogenous opioid peptide genes either decreased or remained unchanged during surgery under intravenous general anaesthesia with epidural block, suggesting that patients remained pain free during surgery.

Establishing the relationship between FGF/MV and FI/FD, and determining the saturation state of sevoflurane anaesthesia during the induction phase and factors affecting nonrespiratory tract ejection volume.

OBJECTIVES: To investigate (i) the relationship between the ratio of fresh gas flow to min volume (FGF/MV) and the ratio of the fraction of inspired sevoflurane to the delivered concentration of sevoflurane (FI/FD); (ii) to establish the saturation state of sevoflurane anaesthesia and factors affecting the volume of ejected sevoflurane through the nonrespiratory tract route (nVERT) during the saturation state. METHODS: Two studies were undertaken in patients with cancer, scheduled to undergo surgery. All patients received tracheal intubation and inhaled sevoflurane. In study 1, anaesthesia parameters were fixed, the initial FD of sevoflurane was 2.2% vol and the FGF/MV was set to 0.2, 0.3, 0.4 and 1.0 in groups A, B, C and D, respectively. In study 2, FGF = MV and the initial FD of sevoflurane was 2.2% vol, but the tidal volume (TV) was set to 6, 8 or 10 ml/kg in groups I, II and III, respectively. RESULTS: Study 1 (n = 60) showed a positive relationship between FI/FD and FGF/MV. In study 2 (n = 60), when TV was fixed, nVERT increased during the saturation state as FD increased; when FD was fixed, nVERT remained stable, despite increased TV. CONCLUSIONS: FI/FD is positively associated with FGF/MV; nVERT is affected by FD but not TV.

Genetic polymorphisms of 15 STR loci in Chinese Han population living in Xi'an city of Shaanxi Province.

Fifteen autosomal STRs loci were analyzed from a population sample of 446 unrelated healthy individuals of Chinese Han population residing in Xi'an city of Shaanxi Province using a multiplex PCR system. We report allele frequencies distribution and statistical parameters for all STR loci, D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818 and FGA. The observed genotype frequencies and expected of genotype frequencies were evaluated by chi(2)-test and the Fisher exact tests. Chi-Square test showed all STR loci were in Hardy-Weinberg equilibrium (p<0.05). Our studied population data were compared with the previously published population data of other ethnics or areas. After comparing, significant differences were found between Chinese Han population in Xi'an and Korean, Tibetan, Uigur, Chinese Han population in Guangdong Province, Hui population, Chinese Salar, Dongxiang and Tu ethnic minority living in Qinghai Province of China at some loci. The forensic parameters from the data showed high values. The data in the present study can be used greatly for routine forensic application in the region of Xi'an city.

Pharmacokinetic and pharmacodynamic population modeling of orally administered rabeprazole in healthy Chinese volunteers by the NONMEM method.

The pharmacokinetic-pharmacodynamic (PK-PD) relationship of the proton pump inhibitor rabeprazole in healthy Chinese volunteers was characterized via a population approach. Healthy Chinese male volunteers were enrolled in the clinical trial. Subjects were divided into three groups by their CYP2C19 genotype. Serum concentrations of rabeprazole were determined using high performance liquid chromatography (HPLC). The intragastric pH values were monitored simultaneously. Data analysis was performed using nonlinear mixed-effects modeling as implemented in the NONMEM software package. The final PK-PD model incorporated a one-compartment PK model with one-order absorption from the gastroenteric trace, first-order elimination pathway with one fixed-effect genotype modeling, and a full sigmoidal Emax PD model (X +/- SE: E0 = 2.30 +/- 0.189; Emax = 7.32 +/- 0.662; EC50 = 51.3 +/- 2.142 ng/ml; Hill coefficient = 5.00 +/- 0.556). The time profiles for concentration and pH value, as well as the concentration-pH value relationship of rabeprazole in healthy Chinese volunteers were well described by the developed population PK-PD model.