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Deconvolution of potential drug targets of the central nervous system (CNS) is particularly challenging because of the complicated structure and function of the brain. Here, a spatiotemporally resolved metabolomics and isotope tracing strategy was proposed and demonstrated to be powerful for deconvoluting and localizing potential targets of CNS drugs by using ambient mass spectrometry imaging. This strategy can map various substances including exogenous drugs, isotopically labeled metabolites, and various types of endogenous metabolites in the brain tissue sections to illustrate their microregional distribution pattern in the brain and locate drug action-related metabolic nodes and pathways. The strategy revealed that the sedative-hypnotic drug candidate YZG-331 was prominently distributed in the pineal gland and entered the thalamus and hypothalamus in relatively small amounts, and can increase glutamate decarboxylase activity to elevate γ-aminobutyric acid (GABA) levels in the hypothalamus, agonize organic cation transporter 3 to release extracellular histamine into peripheral circulation. These findings emphasize the promising capability of spatiotemporally resolved metabolomics and isotope tracing to help elucidate the multiple targets and the mechanisms of action of CNS drugs.
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Objective:To investigate the mechanism of sinomenine (SIN) in inducing the immunosuppressive effects of rat-derived dendritic cells (DCs).Methods:The bone marrow-derived precursor cells from Wistar rats were cultured in vitro. The morphological differences between sinomenine treated DCs (sinomenine modified group, SIN group) and conventional induced DCs (conventional induced group, control group) were observed under microscope. The CD phenotype of DCs was detected by flow cytometry. DCs were induced maturation by lipopolysaccharides (LPS) stimulation. The impact of SIN on the expressions of Toll-like receptor (TLR)2, TLR3 and TLR4 on the DCs surface were detected by flow cytometry. Results:In the conventional induction group, the cells showed clusters or suspension growth, with obvious granular sense on the cell surface; while in the sinomenine induction group, the cells were clustered together, with no significant change in cell volume and morphology. The relative expressions of CD80 and CD86 were 70.7% and 71.3% in the conventional induction group, while 51.7% and 49.4% in the SIN group. The relative expression of TLRs on DCs in SIN + LPS group was TLR2 (51.2±0.34)%, TLR3 (50.3±0.14)%, TLR4 (52.1±0.16)%, which were significantly lower than those in LPS group [(94.35±0.16)%, (97.55±0.16)%, (94.6±0.12)%].Conclusions:SIN may induce immune tolerance by inhibiting the maturation of DCs via inhibiting the TLRs signaling pathways.
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Objective:To summarize the patient profiles and therapeutic efficacies of ABO-incompatible living-related kidney transplantations at 19 domestic transplant centers and provide rationales for clinical application of ABOi-KT.Methods:Clinical cases of ABO-incompatible/compatible kidney transplantation (ABOi-KT/ABOc-KT) from December 2006 to December 2009 were collected. Then, statistical analyses were conducted from the aspects of tissue matching, perioperative managements, complications and survival rates of renal allograft or recipients.Results:Clinical data of 342 ABOi-KT and 779 ABOc-KT indicated that (1) no inter-group differences existed in age, body mass index (BMI), donor-recipient relationship or waiting time of pre-operative dialysis; (2) ABO blood type: blood type O recipients had the longest waiting list and transplantations from blood type A to blood type O accounted for the largest proportion; (3) HLA matching: no statistical significance existed in mismatch rate or positive rate of PRA I/II between two types of surgery; (4) CD20 should be properly used on the basis of different phrases; (5) hemorrhage was a common complication during an early postoperative period and microthrombosis appeared later; (6) no difference existed in postoperative incidence of complications or survival rate of renal allograft and recipients at 1/3/5/10 years between ABOi-KT and ABOc-KT. The acute rejection rate and serum creatinine levels of ABOi-KT recipients were comparable to those of ABOc-KT recipients within 1 year.Conclusions:ABOi-KT is both safe and effective so that it may be applied at all transplant centers as needed.
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Objective To evaluate the effect of the percutaneous nephrolithotomy (PCNL) with the single channel double sets of catheter sheath method in the management of complicated ureteric calculus. Methods The clinical data of forty-eight cases with complicated ureteric calculus were retrospectively investigated. Among them, 18 cases with ipsilateral kidney calculus, 1 case with repeat renal deformity, within the ureteral calculus in upper kidney. Results 48 patients smoothly under ulime from ultrasound guided puncture percutaneous renal channel is established successfully, The establishing the channel to setting the double sheath to the ureteropelvic junction was (18.0 ± 5.0) min, the operation time was (59.0 ± 6.0) min, the stone free rate was 96.6% and no significant complications, such as septic shock, intraoperative or postoperative bleeding and damage to surrounding organs, were detected. The patients were followed up for 3 to 6 months, the degree of hydronephrosis significantly reduced and no ureteral stricture were developed during the period. Conclusion The percutaneous nephrolithotomy (PCNL) with the single channel double sets of catheter sheath method is safe and effective for complicated ureteral calculus.
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Objective To investigate the effect of immature dendritic cell (imDC) derived from donor-derived bone marrow induced by alkaloid sinomenine (SN) on the Th1/Th2 cytokines in venous blood of receptors,and to probe into the mechanism which imDC induced by SN can lead transplantation immune tolerance.Methods Inbred strain Wistar and Sprague Dawley (SD) rats were selected as kidney transplant donor and recipient,respectively.Vessel sutures of the microsurgery technique were used to build the bilaterally renal transplantation model of rats.By the injection of imDC to the recipient rats preoperatively,enzyme-linked immunosorbent assay (ELISA) was used to determine the level of the interleukin (IL)-2,IL-4,IL-10 and interferon-γ (INF-γ).Results (1) Successful rate of transplantation was 89.5％.Arterial anastomosis time was (12.5 ±5.7)min,and venous anastomosis time was (17.3 ± 3.4)min.(2) The content of the IL-2,INF-γ,IL-4 and IL-10 in SN-imDC 106 group was (17.25 ± 3.41) pg/ml,(239.80 ± 9.06) pg/ml,(337.60 ± 25.07) pg/ml,and (1 432.00 ± 106.39) pg/ml,respectively.Among the same concentration,the level of the IL-4 and IL-10 that stood for the Th2 cytokines was significantly higher in SN-imDC group than imDC group and control group (P ＜ 0.05),and was significantly higher in SN-imDC 106 group than SN-imDC 105 group (P ＜ 0.05).Among the same concentration,the lever of IL-2 and INF-γ that stood for the Th1 cytokines was significantly lower in SN-imDC group than imDC group and control group (P ＜ 0.05),and was significantly lower in SN-imDC 106 group than SN-imDC 105 group (P ＜ 0.05).Conclusions (1) The use of microsurgery for anastomosis could make the model of singel kidney transplantation in rats.(2) Specific imDC induced by SN could induce the migration to Th2 immune response,which proved imDC induced by SN could mediate immune tolerance to the recipient.
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Objective To retrospectively analyse the detection results of human leukocyte antigen(HLA) B27,and discuss its diagnosis value in disease such as ankylosing spondylitis(AS).Methods HLA-B27 detection was performed on 1 335 suspected AS patients treated in the hospital from January 2015 to June 2016 by using multiparameter flow cytometry instrument.The test results showed 201 patients were diagnosed with AS(AS group),1 134 patients were diagnosed with rheumatoid arthritis and other related diseases (non AS group).The test results were shown by using mean fluorescence intensity and positive lymphocyte expression rate.n addition,120 healthy people were enrolled as the control group in the physical examination center of the hospital.Results The positive rate of HLA-B27 screening in suspected AS patients was 15.73% (210/1 335),and the positive rate of HLA-B27 screening in the control group was 2.50% (3/120).The positive rate of HLA-B27 screening,the rate of cell expression and the average fluorescence intensity of AS group were 92.03%(185/201),(85.34±17.99)%,8.74±4.20,significantly higher than the non AS group and the control group(P0.05).HLA-B27 screening positive patients were mainly concentrated in adolescence,and the positive rate of male was higher than that of female (P<0.05).Conclusion Flow cytometry examination of HLA-B27 can provide important basis for the diagnosis and differential diagnosis of AS.
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BACKGROUND:It may be an important approach to avoiding organ transplant rejection by utilizing immature dendritic cells to induce donor-specific immunologic tolerance. OBJECTIVE:To study the effect of sinomenine on the differentiation and maturation of rat bone marrow-derived dendritic celsin vitro. METHODS:Bone marrow-derived dendritic cells were isolated from the rat femur and tibia, and immature dendritic cells were induced by granulocyte-macrophage colony stimulating factor and interleukin-4. On day 7, lipopolysaccharide was added and the cells were cultured to generate mature dendritic cells. Cells were divided into control group and low-, middle- and high-dose sinomenine treatment groups (SNL, SNM, SNH groups). Forty hours later, dendritic cels were harvested, and cell morphology was observed by inverted phase contrast microscope. The expression of CD80 and RT1B was detected by flow cytometry. ELISA was used to detect the expression of interleukin-12. The mixed lymphocyte reaction was used to detect the ability of dendritic cells to stimulate the activation of allogeneic T lymphocytes. RESULTS AND CONCLUSION: (1) Under the inverted microscope, the morphology of mature dendritic cells was observed in the control group; in the SNL group most dendritic cells were visible; in the SNM group, there were partially suspended cells with poor maturation; and in the SNH group, most of the cells were not mature. (2) The expression of CD80 in the control group was significantly lower than that in the SNL, SNM and SNH groups (P < 0.05), and the expression of RT1B was significantly reduced in the SNM and SNH groups than the control group. (3) Compared with the control group, the level of IL-12p70 in the cell supernatant was significantly decreased in the SNM and SNH groups (P < 0.01). (4) The ability of dendritic cells to stimulate T lymphocyte proliferation in the SNM and SNH groups was significantly decreased compared with the control group (P < 0.05). To conclude, sinomenine can inhibit the maturation of dendritic cells.
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Objective To analyse the influence of esmolol on plasma N terminal pro-brain natriuretic peptide ( NT-proBNP) , heart-type fatty acid binding protein ( H-FABP) and cardiac function in acute cerebral infarction patients with myocardial enzyme abnormalities.Methods 86 acute cerebral infarction patients with myocardial enzyme abnormalities from September 2010 to July 2014 in our hospital were selected and divided into experimental group and control group randomly (43 cases in each group).All patients were accompanied by myocardial enzyme abnormalities, patients in control group were given basic treatment, while patients in experimental group were given esmolol hydrochloride intravenous injection on the basis of control group.Plasma NT-proBNP, H-FABP levels and cardiac function were analyzed in two groups.Results Myocardial enzyme levels had no significant difference between two groups on admission.Plasma NT-proBNP levels also had no statistical differences after admission hospital of 6 h, 15 h and 3 d , and experimental group was significantly lower than control group ( P<0.05 ) after admission hospital of 5 day and 7 days.H-FABP levels had no statistical differences after admission hospital of 6 h, 15 h, and the experimental group was significantly lower than the control group (P<0.05) after admission hospital of 3, 5, 7 days.Cardiac function with Killip classification showed no significant difference when on admission and after admission hospital of 3 days, and the experimental group was significantly better than control group (P<0.05) after admission hospital of 5, 7 days.Conclusion Acute cerebral infarction patients with myocardial enzyme abnormalities given esmolol on the basic treatments significantly reduce NT-proBNP and HAFBP levels, and improve cardiac function, which is worthy of promotion in clinical applications.
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Objective To investigate the effect and mechanism of hydrogen sulfide (H2S) on kidney injury induced by urinary-derived sepsis. Methods Rabbits were randomly divided into Control group, Sham group, Sepsis group, NaHS 2.8μmol/kg group and NaHS 8.4μmol/kg group. Upper urinary tract obstruction and acute infection was induced to estab-lish Sepsis model. At 24 h before surgery, and 24 h, 48 h, 72 h after surgery, blood was taken to examine white blood cell count (WBC), creatinine (Cr) and blood urea nitrogen (BUN). At 72 h after surgery, morphological changes were observed by HE staining and transmission electron microscopy. Immunohistochemical staining was used to detect TNF-α, IL-10 and NF-κB expression. Blood H2S concentration was measured by deproteinization and Cystathionine-γ-lyase (CSE) activity us-ing spectrophotometric methylene blue method. Results At 24 h, 48 h, and 72 h after surgery, Levels of WBC, Cr and BUN were all elevated in Sepsis group compared with the other four groups. Levels of WBC and BUN in NaHS 8.4μmol/kg group were lower than those in NaHS 2.8μmol/kg group. At 24 h, 48 h after surgery, there is no significant difference be-tween levels of Cr in NaHS 8.4 μmol/kg group and that in NaHS 2.8 μmol/kg group , but Cr level in NaHS 8.4 μmol/kg group was marked lower than that in NaHS 2.8μmol/kg group 72 hour after surgery. Pathological features of kidney injury were also alleviated by intravenous administration of NaHS. TNF-α, NF-κB expressions in NaHS 2.8μmol/kg group and NaHS 8.4 μmol/kg group were lower than those in Sepsis group, IL- 10 expression was higher than that in Sepsis group. TNF-α, NF-κB expressions in NaHS 8.4μmol/kg group were lower than that in NaHS 2.8μmol/kg group, whereas IL-10 expression in NaHS 8.4μmol/kg group was higher than that in NaHS 2.8μmol/kg group. Compared with Control group and Sham group, H2S content and CSE expression in kidney were decreased in Sepsis group. After intravenous administration of NaHS, H2S content increased, but the CSE activity has no obvious change. Conclusion Exogenous H2S reduced kidney in-jury induced by urinary-derived sepsis through inhibiting NF-κB, decreasing TNF-αand increasing IL-10.
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Objective To investigate the effect of gene silencing of Y-box binding protein-1(YB-1) by RNA interference on the proliferation and migration in prostate cancer cells lines PC-3 cells .Methods YB-1 siRNAs(pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2) were synthesized and transfected into cloned into the the PC-3 cells by liposome .The expressions of YB-1 were measure by RT-PCR and Western blotting .The proliferation and migration were respectively detected by MTT and Transwell method . Results ThemRNA and protein expressions of YB-1 were significantly decreased by pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2 (P<0 .05) ,compared with the control group ,the inhibition ratio of mRNA expression was 36 .23% and 39 .42% respectively and the inhibition ratio of protein expression was 41 .56% and 55 .33% respectively .The proliferation and migration were significantly decreased by pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2(P<0 .05) .Conclusion YB-1 gene silencing by RNA interference inhibits the proliferation and migration in prostate cancer cells lines PC-3 cells .
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Objective To observe the efficacy of CTLA4Ig and CD40Ig gene local cotransfection on the survival of renal allografts.Methods The kidneys of guinea pig were transfected with PcDNA 3.1+-CTLA4Ig and/or PcDNA 3.1+ -CD40Ig gene by Lipo2000,and the transfected kidneys were transplanted to SD rats.The recipients were divided into group 1 (transfected with PcDNA 3.1 +,mock group),group 2 (transfected with PcDNA 3.1 + -CD40Ig,CD40Ig group),group 3 (transfected with PcDNA3.1+ -CTLA4Ig,CTLA4Ig group) and group 4 (co-transfected with PcDNA 3.1+-CD40Ig and CTLA4Ig,CTLA4Ig+ CD40Ig group).The effects of CTLA4Ig and CD40Ig transfection were determined by Western blotting.The serum creatinine (Scr) in the recipients,the pathological changes of the allografts and the survival of renal allografts were observed.Results Significantly prolonged allografts survival time was observed in group 2 (40.7 ± 10.9 days),group 3 (49.3 ± 9.5 days) and group 4 (75.7 ± 8.0 days) as compared with group 1 (6.8 ± 1.9days),especially allografts got the longest survival time in group 4 (75.7± 8.0 days).The serum creatinine level was reduced in group 4 at 30th day as compared with that in group 2 and group 3.The lymphocytes infiltrating rate in the grafts was lowest in group 4 after transplantation.Conclusion Local co-transfection of CTLA4Ig and CD40Ig genes can prolong the survival time of renal allografts significantly.
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Objective To investigate the expression of CD40 protein in bladder urothelium cancer and to explore its correlation with clinical stage, pathological grades, cell apoptosis. Methods Expression of CD4o protein was evaluated by immunohistochemistry in paracancerous tissues (20 cases) and bladder urothelium cancer tissues (78 cases). Apoptosis of bladder urothelium cancer tissues was detected by Hoechst to determine the apoptotic rate (AR). Results The expression rate of CD40 protein was 2/20(10 %) in the cases of paracancerous tissues, 55/78(70.5%) in the cases of bladder urothelium cancer tissues, a statistically significant differences of CD40 staining was observed between paracancerous tissues and bladder urothelium cancer tissues (P ＜0.01). Overexpression of CD40 was strongly inversely related to the tumor grade and clinical stage; the positive group of CD40 protein was (12.60±0.38)%, the negative group of CD40 protein was (6.77±0.53)%, a statistically significant differences of apoptosis were observed between CD40 protein positive group and CD40 protein negative group (P ＜0.01). It showed CD40 protein might induce cell apoptosis. Conclusion Expression of CD40 protein is closely related with clinical stage, pathological grade and cell apoptotic rate and may offer reliable evidence for diagnosis, treatment and prognosis in bladder urothelium cancer.
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Objective To study the immunosuppressive mechanism of alkaloid sinomenine (SIN) by observing the effects of SIN on the proliferation and intracellular protein expression levels of nuclear factor of activated T cells (NF-AT) and interferon-gamma (IFN-γ) in CD4+ T lymphocytes of human periphery blood. Methods CD4+ T lymphocytes were isolated from PBMC suspensions with immunomagnetic beads and divided into five groups to culture. (1) Negative control group: no medicine was added to cell culture medium; (2) Positive control group: CsA solution (final concentration: 50ng/ml) was added to cell culture media; (3) Low-concentration SIN group (L-SIN): low-concentration SIN solution (final concentration: 10 μmol/L) was added to cell culture media; (4) Middle-concentration SIN group (M-SIN): middle-concentration SIN solution (final concentration: 200 μmol/L) was added to cell culture media; (5) High-concentration SIN group (H-SIN): high-concentration SIN solution (final concentration: 1000 tmol/L) was added to cell culture media. The proliferations of CD4+ T lymphocytes were observed. Western blotting was performed to detect the protein expression levels of NF-AT. FCM was used to determine the levels of IFN-γ. Results Compared with negative control group, the cell proliferation was significantly inhibited in H- and M-SIN groups (P＜0. 01 ). SIN concentration-dependently inhibited the protein expression levels of NF-AT and IFN-γ in CD4+ T lymphocytes of human periphery blood (P＜0.01). The protein expression levels of NF-AT and IFN-γ were lowest in positive control group. There was a close negative correlation between intracellular levels of NF-AT and cell proliferation inhibition ratio in CD4+ T lymphocytes of human periphery blood (rs = - 0. 969, P = 0. 000). Conclusion SIN can inhibit the protein expression of NF-AT and IFN-γ in CD4+ T lymphocytes of human periphery blood probably by decreasing protein levels of NF-AT to inhibit the activity and proliferation of CD4+ T lymphocytes.
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Prediction of RNA secondary structures including pseudoknots is a difficult topic in RNA field. Current predicting methods usually have relatively low accuracy and high complexity. Considering that the stacking of adjacent base pairs is a common feature of RNA secondary structure, here we present a method for predicting pseudoknots based on covariance with stacking and minimum free energy. A new score scheme, which combined stacked covariance with free energy, was used to assess the evaluation of base pair in our method. Based on this score scheme, we utilized an iterative procedure to compute the optimized RNA secondary structure with minimum score approximately. In each interaction, helix of high covariance and low free energy was selected until the sequences didn't form helix, so two crossing helixes which were selected from different iterations could form a pseudoknot. We test our method on data sets of ClustalW alignments and structural alignments downloaded from RNA databases. Experimental results show that our method can correctly predict the major portion of pseudoknots. Our method has both higher average sensitivity and specificity than the reference algorithms, and performs much better for structural alignments than for ClustalW alignments. Finally, we discuss the influence on the performance by the factor of covariance weight, and conclude that the best performance is achieved when lambda1 : lambda2 = 5 : 1.
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Algorithms , Base Pairing , Base Sequence , Computational Biology , Methods , Molecular Sequence Data , Nucleic Acid Conformation , RNA , Chemistry , Genetics , Sequence Analysis, RNAABSTRACT
The nature and preparation of sodium alginate are introduced and its medical applications are reviewed in fields such as medical carrier,gel microspheres,sustained-release materials,surgical wound repairing materials,and so on.Besides,its development prospects are described.It is an inevitable trend that sodium alginate is used in the development of controlled-release reagent.There are also important applications in the partial treatment of ear disease and artificial cartilage for sodium alginate,but the strength and toughness of alginate hydrogel limits its wider application.
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The teaching secretary of clinical department at medical university is required to be passionate,devoted,qualified and good at writing.The position is reserved for the person with healthy psychology,who knows well to deal with people,political training and sensitivity,advanced qualification.
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Objective To afford the basis for the application of Sinomenine(SIN) in the proposal for organ transplantation, the expression of TGF-? 1 was explored in inborn WistarSK kidney transplantation model. Methods Forty-eight recipients were divided into 4 groups at random:group 1 being used as control and treated with 0 9% natrii chloride(3ml?kg -1 ?d -1 ,ip), group 2 treated with SIN(30mg?kg -1 ?d -1 ,ip),group 3 treated with CsA(2 5mg?kg -1 ?d -1 ,ip),group 4 treated with SIN(30mg?kg -1 ?d -1 ,ip) combined with CsA(2 5mg?kg -1 ?d -1 ,ip).Allografts of 6 recipients from each group were harvested at day 6 before end stage rejection for histological studies. The synergistic effects of SIN combined with CsA were assessed by COX proportional hazards model. The quantitative methods of immunohistochemistry were used for analysis on expression of TGF-? 1.Results Control recipients rejected their allografs at day 9 post-operation, the average life time was 7 83?9 75 days. Treated with either SIN(30mg?kg -1 ?d -1 ,ip) or subtherapeutic doses of CsA(2 5mg?kg -1 ?d -1 ,ip) allografts survival was mininally prolonged. Combined treatment with SIN and subtherapeutic doses of CsA led to a significant prolongation of graft survival(group 4), more than 18 days. The expression of TGF-? 1 was enhanced significantly in group 2 and group 3.No synergistic effect of SIN combined with CsA on expression of TGF-? 1 was observed. Conclusion The therapeutic value of SIN in rat kidney allograft model suggests that this safe chinese medicine has remarkable efficacy of anti-acute rejection in rat kidney transplantation. It shows significant synergistic effect with sub-therapeutic CsA. The immunosuppressive efficacy of SIN is probably based on enhancing express of TGF-? 1. No synergistic effect of SIN and CsA on expression of TGF-? 1 indicates low danger of fibrosis of allografts. SIN will be a hopeful chinese medicine for immunosuppressive treatment in organ transplantation.
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0.999 0) and higher precision (RSD