The paper is to report the study of pharmacokinetics of transdermal administered nicotine in the brain of freely moving rat by using microdialysis with stable labeled isotope as internal standard. The pharmacokinetic behavior of nicotine in Sprague Dawley rat brain was investigated after intranasal administration (3.75 mg). Brain fluid samples were collected by intracerebral microdialysis with DL-nicotine as internal standard. Concentrations of nicotine and DL-nicotine in the sample were measured by HPLC-MS/MS. Main pharmacokinetic parameters were calculated and analyzed by Das 2.0 pharmacokinetic software. The recovery of nicotine and the delivery of DL-nicotine were the same. The fate of absorption and distribution was two compartment model and the values of t1/2alpha was 170.31 min, t1/2beta was 263.30 min and the AUC(0-infinity) was 2.75 x 10(5) microg x L(-1) min separately. DL-nicotine can be used to calibrate the recovery of nicotine, and the new method of stable isotope microdialysis can be used to study the pharmacokinetics of freely moving rat. It will make sense for the treatment of addiction of tobacco and provide a new thought for the research of pharmacokinetics-pharmacodynamic combination.

Objective To investigate the impacts of γ-ray radiation on gene expressions of peripheral blood lymphocytes in SD rats,and to screen differential expression genes and biological pathways closely related to radiation injury.Methods The differential gene expression of peripheral blood lymphocytes in SD rats was selected by microarray at 6 h after 2 Gy 60Co γ-ray exposure in vitro and in vivo.Bioinformatics analysis of the differentially expressed genes was performed by using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases.Real-time quantitative PCR was applied to verify the screen results of the microarray.Results Fifty-five genes with three times over-expressed level were screened out from the radiation groups both in vitro and in vivo and they were involved in 6 signaling pathways.There were two differentially expressed genes of microarray assay were verified by PCR assay.Conclusions Differential expressed genes in the peripheral blood lymphocytes of SD rats could be induced by γ-ray radiation and they cooperatively contributed to a variety of biological processes.

Objective To investigate the changes of gene expression profile in Beagle dogs' peripheral blood lymphocytes after acute irradiation.Methods Totally 20 male adult Beagle dogs were randomly divided into four groups including non-treatment blank control group and three radiation groups exposed to 0.5,2.0,and 5.0 Gy of γ-rays,respectively.Six hours after radiation,the peripheral blood were collected for lymphocytes isolation.Total RNA was extracted from the lymphocytes and analyzed by microarray hybridization.The differential gene profiles of radiation groups were analyzed by gene ontology (GO) analysis and kyoto encyclopedia of genes and genomes (KEGG) pathways analysis,and the alerted genes were further confirmed by the real time quantitative PCR (qRT-PCR) assay.Results Compared to the blank control group,the expressions of 308 genes in the radiation groups were perturbed over 2-fold of control,including 61 genes up-regulated and 247 genes down-regulated,which were mainly associated with immune response and cancer occurrence.The GO analysis indicated that the differential expressed genes were associated with cell connection,signal transduction,oxidation-reduction reaction and metabolism.The KEGG pathway analysis showed that some physiological and biochemical processes were involved,such as phagocytosis,tumor pathway,p53 signaling pathway,JAK-STAT signal pathway,cell differentiation,and proliferation,oxidative phosphorylation,glycogen dysplasia and other pathways.The microarray data of the alterations of apoptosis enhancing nuclease (AEN) and mitogenactivated protein kinase 13 (MAP3K13) gene expressions were further confirmed by qRT-PCR.Conclusions Exposure to different doses of acute γ-ray irradiation had significant impact on the gene expressions in Beagle dogs' peripheral blood lymphocytes and those differential expressed genes were related to a series of biological processes and pathways including immune response,metabolism and carcinogenesis.

Objective To observe the protective effects of different doses of hydrogen-rich water on radiation injury in mice,so as to provide scientific basis for the application of hydrogen-rich water.Methods The ICR mice were randomly divided into control group,irradiation group,amifostine group and hydrogen-rich water of low,medium and high dose groups.The 30 days survival rate,body weight,hematology parameters,serum biochemical parameters,organ weight and coefficient,bone marrow micronucleus rate,bone marrow nucleated cell count were observed after total body irradiation with 9.0 Gy gamma rays.Results After 30 d of irradiation,the hydrogen-rich water showed obvious protective effect on the survival rate and body weight in a dose dependent manner so that the survival was significantly higher than that of irradiation group (t =-2.67,P ＜ 0.05).The biochemical index,such as TP,ALB and CRE in the low dose group,TP,ALB,TBIL and CRE in the medium dose group,and TP,ALB,GLU,TBIL,BUN,GRE and UA in the high dose group also indicated the protective effects of hydrogen-rich water (t =-2.04--4.11,P ＜ 0.05).But the protective effect of hydrogen-rich water was not observed in hematology,organ weight and coefficient,and bone marrow micronucleus induction.Conclusions The hydrogen-rich water has anti-radiation effect,which may depend on the dose of hydrogen.

Objective To construct lentiviral vectors containing peptide P1-GFP fusion genes.Umbilical cord derived mesenchymal stem cells were infected with lentivirus carrying peptide P1 and GFP fusion genes.To inject the targeted umbilical cord derived mesenchymal stem cells into mice and to detect GFP expression in the spleen.Methods Umbilical cord derived mesenchymal stem cells were cultured with adhered tissues of umbilical cord smaller than 1 mm3 . Lentiviral vector containing P1-GFP fusion genes with engineering technology was constructed and infected the umbilical cord derive mesenchymal stem cells.Targeted umbilical cord derived mesenchymal stem cells were intravenously injected in the mouse tail vein and after 18 hours GFP expression was detected with immunohistochamical staining of the spleen tissues.Results Harvested umbilical cord derived mesenchymal stem cells grew well in culture medium. Green fluorescence on umbilical cord derived mesenchymal stem cells were observed under fluorescence microscope at 18 hours after infected with lentivirus.Green fluorescence intensity of umbilical cord derived mesenchymal stem cells was increasing over time and reached a peak at 72 hours.Umbilical cord derived mesenchymal stem cells highly expressed CD105 (90.0%)/CD44 (98%) and CD73 (85.0%)/CD90 (98.5%) molecules.GFP expression was detected in the spleen after intravenous injection of targeted umbilical cord derived mesenchymal stem cells in the mice 18 hours later.GFP expressing cells intimately contacted with lymphocytes.Conclusions Targeted umbilical cord derived mesenchymal stem cells contain P1-GFP fusion genes are constructed.Targeted umbilical cord derived mesenchymal stem cells can be targeted to mouse spleen and intimately contact with lymphocytes after intravenous injection.Our results lay the groundwork for further studies.

Objective:To explore the intervention effect of Shumu Peitu herb-partitioned moxibustion on clinical signs and symptoms and negative emotions of diarrhea-predominant irritable bowel syndrome (IBS-D) patients with liver-stagnation and spleen-deficiency pattern.Methods:A total of 72 patients with IBS-D of liver-stagnation and spleen-deficiency pattern treated in the Department of Gastroenterology of Nanjing Vniversity of Chinese Medicine from September 2021 to June 2022 were selected for randomized controlled trial. The patients were randomly divided into the observation group (2 cases dropped off, 34 cases in total) and control group (1 case dropped off, 35 cases in total) by random number table method. The patients in control group were treated with Tongxieyaofang (TXYF). The patients in observation group were treated with oral administration of TXYF and Shumu Peitu herb-partitioned moxibustion, and both groups were treated for 4 weeks. The clinical efficacy, Traditional Chinese Medicine (TCM) syndrome integral, IBS Quality of Life Questionnaire (IBS-QOL), IBS Symptom Severity Scale (IBS-SSS), Bristol Stool Form Scale and Hospital Anxiety and Depression Scale (HADS) were compared before and after treatment.Results:After treatment, the total effective rate of the observation group was 94.12%(32/34), which was higher than the 71.43%(25/35) in the control group, the difference was significant ( χ2 = 6.18, P<0.05). After treatment, the TCM syndrome integral in the observation group was (7.62 ± 4.08), which was lower than the (9.89 ± 4.71) in the control group, the difference was significant ( t = 2.14, P<0.05). After treatment of 3 days, the scores of quality of life in the five dimensions of dysthymia, behavior disorder, health worry, avoidance of eating and social function in the observation group were (82.44 ± 11.46), (80.25 ± 11.67), (76.23 ± 12.67), (59.80 ± 15.14) and (79.23 ± 11.59) points, which were different with the (73.57 ± 12.39), (72.35 ± 15.48), (69.76 ± 13.11), (50.00 ± 16.17) and (73.04 ± 13.11) points in the control group, the difference were significant ( t values were -3.09 - -2.08, all P<0.05). Three days after treatment, the score of IBS-SSS and Bristol fecal character in the observation group were (118.24 ± 40.64) and (5.09 ± 0.62) points, which were lower than the (146.86 ± 60.09) and (5.51 ± 0.66) points in the control group, the difference were significant ( t = 2.31 and 2.76, both P<0.05). After treatment, the score of HADS-A and HADS-D in the observation group were (6.26 ± 1.75) and (5.29 ± 1.47), which were different with the (7.26 ± 2.19) and (6.17 ± 2.11) in the control group, the difference were significant ( t = 2.08 and 2.00, both P<0.05). Conclusions:Shumu Peitu herb-partitioned moxibustion can effectively improve IBS-D patients with liver-stagnation and spleen-deficiency pattern, relieve clinical symptoms, reduce negative emotions, and improve quality of life.

OBJECTIVETo investigate the effect of aluminum exposure on neuronal apoptosis of rats hippocampus and the correlation of and synaptic plasticity.

METHODSThere were 40 SPF grade SD rats which were randomly divided into four groups: the control group, the low dose group, the medium dose group and the high dose group, 10 rats in each group. The rats were daily gavaged with aluminum lactate for 30 days. The hippocampal fEPSPs in rat was measured by electrophysiological grapher and the neuronal apoptosis in hippocampus was detected by Flow cytometer. In addition, the relative expression of gene which includes caspase-3, 8, 9 was measured by Real-time PCR.

RESULTSCompared to the control group, the average of fEPSPs which after HFS 10, 20, 30, 40, 50, 60 min was decreased at different time point in the low dose group, the medium dose group and the high dose group (P < 0.05). Compared with the control group, the rate of apoptosis was significantly increased in the medium dose group and the high dose group (P < 0.05). Compared to the control group, the relative expression of caspase-3 in the medium dose group and the high dose group was significantly increased in Real-time PCR (P < 0.05), and the relative expression of caspase-8 in the high dose group was significantly increased (P < 0.05).

CONCLUSIONAluminum exposure may induced neuronal apoptosis in rats, and then affect hippocampal synaptic plasticity.

Aluminum ; toxicity ; Aluminum Compounds ; toxicity ; Animals ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Hippocampus ; cytology ; drug effects ; Lactates ; toxicity ; Neuronal Plasticity ; drug effects ; Neurons ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley

Objective To investigate the protective effect of hydrogen-rich water on rat cognitive dysfunction induced by ionizing radiation.Methods A total of 20 SD rats were randomly divided into four groups with the ramdom number table method: control group(C),hydrogen-rich water group(HRW),irradiation group(IR)and hydrogen-rich water intervention group(HRW+IR),with 5 rats in each group.The spatial memory ability of rats was tested by a morris water maze.The expression of apoptosis-related genes was detected by real-time fluorescence quantitative PCR.The changes of glutathione(GSH),8-hydroxydeoxy guanosine(8-OHdG)and malondialdehyde(MDA)and SOD were also measured.Results The escape latency(F＝6.003,P＜0.05)and the swimming distances(F＝3.850,P＜0.05)of rats in four different groups had statistically significant differences.Compared with the IR group,the escape latency of the HRW+IR group was significantly decreased at 3,4,5 d after irradiation(P＜0.05),and the swimming distance of this group became much longer(P＜0.05).The levels of GSH,8-OHdG and MDA in these four groups had statistically significant differences(F＝6.450,5.033,4.113,P＜0.05).Compared with IR group,the concentration of GSH was increased(P＜0.05),but MDA and 8-OHdG decreased(P＜0.05)in the brain tissue of HRW+IR group,and the expressions of caspase-3,caspase-9 and bax genes were reduced(t＝2.956,3.087,5.246,P＜0.05),while the expression of bcl-2 gene was enhanced(t ＝-3.640,P ＜0.05)in the HRW+IR group.Conclusions Hydrogen-rich water attenuates the oxidative damage of ionizing radiation by neutralizing oxyhydrogen free radicals and thus protects brain from radiation damage.

Objective To investigate the endocytosis and exocytosis of soluble uranium in human kidney proximal tubular epithelial (HK-2) cells and the cytotoxicity after uranium exposure. Methods Cell Counting Kit-8 assay was used to determine the cell viability after different concentrations of uranium exposure, and optical microscopy and transmission electron microscopy were used to observe the changes in cells after uranium exposure. Inductively coupled plasma mass spectrometry was used to monitor the endocytosis and exocytosis of uranium over time by cells. Flow cytometry was used to assess the changes in cell cycle and apoptosis after uranium exposure. Results After uranium exposure, HK-2 cells showed dose-dependent damage; cell cycle was arrested in G1 phase; cell apoptosis and necrosis occurred; cell proliferation was inhibited. The content of endocytic uranium increased gradually within 24 h, and there was a threshold for uranium endocytosis, while the fraction of uranium binding to cell surface was low (< 0.2%). Over 40% of the endocytic uranium would be exocytosed within 1 h. Uranium could form needle-like precipitates in both intracellular and extracellular areas after uranium exposure. Conclusion After uranium exposure, cells show decreased viability, cell cycle arrest, and cell apoptosis. The process of endocytosis and exocytosis of soluble uranium is very rapid. HK-2 cells can convert soluble uranium into non-toxic precipitates.

Objective: To establish a method for the determination of aluminum in blood, and to detect the aluminum content in the blood of occupational aluminum workers. Methods: The morning blood of the aluminum workers was collected in an anticoagulation tube, and the supernatant was centrifuged. The supernatant was diluted with 4% nitric acid containing 1% Triton for 24 h at room temperature, and the supernatant was centrifuged. The supernatant was filtered and the blood aluminum concentration was measured by inductively coupled plasma mass spectrometry and quantified by external standard method. Results: The detection limit of ICP-MS method was 0.39 μg/L, the linear range was 0-160 μg/L, the recoveries were 98.24%-99.65%, and the precision was 0.19%-0.28%. The recoveries of graphite furnace atomic absorption spectrophotometry (GFAAS) were 97.17%-111.18%, and the precision was 0.35%-0.44%. The average blood aluminum concentration of aluminum workers in the normal control group was (19.87±10.65) μg/L. The average blood aluminum concentration of aluminum workers in the expose group was (31.12±11.43) μg/L. Conclusion The method of ICP-MS for the determination of aluminum concentration in blood has a simple pretreatment process, high recovery rate, low detection limit and high precision, which is suitable for popularization.