Traditional Chinese medicine has a long history of intranasal administration. Compared with the other administration routes, intranasal administration has the benefits of fast absorption, high bioavailability, high brain-targeting and non-invasive. In the past few years we take "Xingnaojing" and "Tongqiao Sanyu formula" as model drug and studied pharmacokinetics of effective components of different polarities. MDCK/MDCK-MDR1 cells were used to simulate blood brain barrier to study the permeate behaviors of different drug and the mechanism of enhancing effects of aromatic medicine. Then a microemulsion (modified by mPEG2000-PLA) was prepared for intranasal administration, and the pharmacokinetics and investigated tissue distribution were studied by fluorescence imaging. The irritation of the drug and different preparations were studied on human nasal epithelial cell (HNEC) cell and living animals. In this paper, we reviewed the achievements and hope that it can provide constructive suggestions for the future research.
Administration, Intranasal ; instrumentation ; methods ; Animals ; Biological Availability ; Blood-Brain Barrier ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Humans

Objective To evaluate the clinical diagnostic value of prospective electrocardiograph (ECG)-gating of CT cardiac angiography in congenital heart diseases of Chinese population through a Meta-analysis.Methods The articles were searched to study CT prospective ECG-gating in diagnosis of congenital heart disease from January 1995 to February 2016 in domestic and foreign publications.The study quality was assessed by the Quality Assessment for Diagnostic Accuracy Studies and the data extraction was performed.The software of Meta-disc1.4 was used for heterogeneity test.Different effect models were choosen according to the results of heterogeneity analysis.Meanwhile,this soft was used to calculate the sensitivity,specificity,likelihood ratio and its 95％ confidence interval (CI),respectively.The forest maps and summary receiver operator characteristic (SROC) curve were drawn.In addition,the area under curve (AUC) was calculated.Results Twelve articles were included in the Meta-analysis.The study included 1 431 congenital heart malformations confirmed by surgery or cardiac catheterization angiography.CT prospective ECG-gating technique had no heterogeneity in sensitivity and specificity of congenital heart disease.The total sensitivity,the total specificity,positive likelihood ratios,negative likelihood ratios and its 95％ CI of CT cardiac angiography were 96％ (95％ CI 94％ to 97％),100％ (95％ CI 100％ to 100％),365.94(95％ CI 231.18 to 579.26),0.04(95％ CI 0.03 to 0.05) with fixed effect model,respectively.The AUC of the SROC was 99.86％,Q =0.987 9.Conclusions Prospectively ECG-gating of CT cardiac angiography has high sensitivity and specificity in the diagnosis of congenital heart diseases.Its AUC of the SROC is large.It has high diagnostic value in congenital heart diseases.

In order to investigate the characteristics of transdermal delivery of ferulic acid under the treated of microneedle arrays and the influence on permeability of rat skin capillaries, improved Franz-cells were used in the transdermal delivery experiment with the rat skin of abdominal wall and the length of microneedle arrays, different insertion forces, retention time were studied in the influence of characteristics of transdermal delivery of FA. The amount of FA was determined by HPLC system. Intravenous injection Evans blue and FA was added after microneedle arrays treated. Established inflammation model was built by daubing dimethylbenzene. The amount of Evans blue in the rat skin was read at 590 nm wavelength with a Multiskan Go microplate reader. Compared with passive diffusion group the skin pretreated with microneedle arrays had a remarkable enhancement of FA transport (P <0.01). The accumulation of FA increased with the enhancement of insertion force as to as the increase of retention time. Microneedle arrays with different length had a remarkable enhancement of FA transport, but was not related to the increase of the length. The research of FA on the reduce of permeability of rat skin capillaries indicated that the skin pretreated with microneedle arrays could reduce the content of Evans blue in the skins of rat significantly compared with the untreated group. The permeation rate of ferulic acid transdermal delivery had remarkable increase under the treated of microneedle arrays and the length of microneedle arrays ,the retention time so as to the insertion force were important to the transdermal delivery of ferulic acid.
Administration, Cutaneous ; Animals ; Coumaric Acids ; administration & dosage ; pharmacokinetics ; Male ; Needles ; Rats ; Rats, Sprague-Dawley ; Skin Absorption

Objective To explore the relationship between blood glucose and lipid levels and cognitive function in elderly patients with comorbidity of depression and type 2 diabetes mellitus (T2DM). Methods The clinical data from elderly participants (60 to 79 years old) receiving physical examination between Nov. 1 and Dec. 30, 2017 were collected. According to inclusion and exclusion criteria, 59 cases with comorbidity of depression and T2DM were assigned to comorbid group, 106 depression cases were in depression group, 84 T2DM cases were in diabetes group and 248 were in control group (with no diabetes or depression). The general physiological indicators (height, body mass, waist circumference, hip circumference and blood pressure) were collected, the body mass index (BMI) and waist-to-hip ratio (WHR) were calculated, and blood glucose and lipid levels were determined. The Montreal Cognitive Assessment (MoCA) scale was used to assess the cognitive function of the elderly in each group. The differences in BMI, WHR, blood pressure, blood glucose level and blood lipid level among the groups were compared, and the relationships between these indicators and the adjusted total score and scores in each cognitive domain of MoCA scale were analyzed. Results (1) There were no significant differences in height, body mass, BMI, WHR or diastolic blood pressure (DBP) among four groups (all P>0.05), while the differences in systolic blood pressure (SBP) and pulse pressure (PP) were significant (both P<0.01), with the increase in the diabetes group being most obvious. (2) Compared with the control group, the fasting blood glucose (FBG) level, oral glucose tolerance test (OGTT) 2 h and glycosylated hemoglobin (HbA1c) level were significantly higher in the comorbid group and the diabetes group (all P<0.01); while there was no significant difference between the control and the depression groups (P>0.05). The triglyceride (TG) level in the comorbid group was significantly higher than that in the control group (P<0.05), and the high-density lipoprotein (HDL) levels in the comorbidity and the diabetes groups were lower than that in the control group (P<0.05, P<0.01). (3) There was no significant difference in the adjusted total score of MoCA scale among the four groups (P>0.05). Compared with the control group, the attention scores of the other three groups were significantly lower (all P<0.01). The elderly in the comorbid group had significantly lower fluency and orientation scores compared with the elderly in the control group (P<0.05, P<0.01), and had significantly lower orientation score compared with the elderly in the diabetes group (P<0.05). (4) Simple linear regression analysis showed that the adjusted total score of MoCA scale was negatively correlated with FBG and HbA1c levels (b=-0.339, P=0.006; b=-0.482, P=0.023), and the attention score was negatively correlated with FBG, OGTT 2h and HbA1c levels (b=-0.119, P<0.001; b=-0.040, P=0.002; b=-0.161, P=0.006). (5) Multiple linear regression analysis revealed that FBG level was negatively correlated with the adjusted total score of MoCA scale (B=-0.349, P=0.004). Conclusion Hyperglycemia may be a risk factor of cognitive dysfunction of elderly patients with comorbid of depression and T2DM.

Objective To observe the sequence of fat deposit and its relationship with insulin resistance in SD rats fed by high fat diet.Methods Normal 8-week-old male SD rats were randomly divided into normal chow (NC,n=40)and high fat diet(HF,n=40)groups.Triglyceride(TG)in serum,liver and muscle were measured;glucose infusion rate(GIR)and the mRNA level of genes related to lipid metabolism in liver and muscle were determined in different stages.GIR was detected by eugiyeemic-hyperinsulinemic clamp for evaluating the insulin sensitivity.Gene expression was determined by real-time PCR.Results(1)As compared with NC group,serum TG was not increased after high fat feeding for4 and 8 weeks,it began to increase after 12 weeks [0.52(0.15-1.00) mmol/L vs O.31(0.09-0.53)retool/L, P0.05)in skeletal muscle.After 8 weeks,the expression of ACC1 in liver in HF group was increased by 20.6%,CPT-1 was decreased by 27.1%(P

OBJECTIVEPulmonary vascular structural remodeling induced by high pulmonary blood flow is an important pathologic basis of pulmonary hypertension with congenital heart disease of left-to-right shunt. However, the mechanism is still not clear. The present study aimed to examine the alteration of endogenous nitric oxide (NO) pathway in high pulmonary blood flow-induced pulmonary vascular structural remodeling, so as to explore the role of NO pathway in pulmonary hypertension induced by high pulmonary blood flow.

METHODSSixteen male SD rats were randomly divided into control group (n = 8) and shunting group (n = 8). Aortocaval shunting was produced for 11 weeks in shunt rats. Pulmonary artery mean pressure (mPAP) of each rat was evaluated using right cardiac catheterization. The ratio of right ventricular mass to left ventricular plus septal mass [RV/(LV + S)] was detected. Pulmonary vascular micro-and ultra-structure was examined by using a light microscope and a transmitted electronic microscope. Meanwhile, the concentration of plasma NO was measured by spectrophotometry. The expressions of endothelial NO synthase (eNOS) mRNA and protein by pulmonary arteries were detected by in situ hybridization and immunohistochemistry, respectively.

RESULTSAfter 11-week aortocaval shunting, mPAP was significantly increased [(22.5 +/- 2.6) mmHg vs. (15.8 +/- 2.8) mmHg, 1 mmHg = 0.133 kPa, t = 4.97, P < 0.01], and RV/(LV + S) was also markedly increased (0.267 +/- 0.022 vs. 0.221 +/- 0.016, t = 4.85, P < 0.01). The percentage of muscularized arteries was obviously increased in shunt rats compared with controls [(23.2 +/- 2.4)% vs. (13.5 +/- 2.1)%, t = 7.82, P < 0.01], and relative medial thickness of pulmonary arteries was obviously increased in shunt rats [median pulmonary artery: (7.76 +/- 0.56)% vs. (4.82 +/- 1.03)%, t = 6.23, P < 0.01; small pulmonary artery: (11.94 +/- 0.66)% vs. (6.91 +/- 0.53)%, t = 14.96, P < 0.01]. Ultrastructural changes, such as hyperplasia and degeneration of endothelial cells, irregularity of internal elastic laminar and hypertrophy and the increased number of synthetic phenotype of smooth muscle cells, were found in intrapulmonary arteries of shunt rats. Meanwhile, plasma NO concentration was increased [(30.2 +/- 7.9) micromol/L vs (19.7 +/- 5.7) micromol/L, t = 3.05, P < 0.01) and eNOS mRNA and protein expressions by pulmonary arteries were significantly augmented in rats of shunting group.

CONCLUSIONThe upregulation of eNOS/NO might be an adaptive response of pulmonary circulation to an increased blood flow in the development of pulmonary hypertension and pulmonary vascular structural remodeling.

Animals ; Blood Flow Velocity ; Hypertension, Pulmonary ; physiopathology ; Immunohistochemistry ; In Situ Hybridization ; Male ; Nitric Oxide ; blood ; Nitric Oxide Synthase ; blood ; genetics ; Nitric Oxide Synthase Type III ; Pulmonary Artery ; physiopathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo determine the O/W partition coefficient of panax pseudo-ginseng saponin (PNS) and investigate the absortion kinetics of it in whole small intestine and different intestinal segments of rats.

METHODThe shake-flask method was employed to determine the O/W partition co-efficient of geniposide, and an in situ intestinal perfusion model was employed to investigate the absorptive kinetics of geniposide.

RESULTThe partition coefficient (P) of R1, Rg1 and Rb, of PNS were 1.0814, 6.3104 and 0.2743, respectively, and their logP were 0.0340, 0.8001, -0.5618, the absorptive rate constants (Ka) of R1, Rg1 and Rb1 of PNS at the concentration of 0.2, 0.5, 1.0 g x L(-1) were (0.135 +/- 0.006), (0.110 +/- 0.002), (0.095 +/- 0.016), (0.144 +/- 0.015), (0.110 +/- 0.006), (0.099 +/- 0.011), (0.238 +/- 0.013), (0.140 +/- 0.008), (0.137 +/- 0.012)h(-1), respectively. The Kb of R1, Rg1 and Rb1 of PNS were (0.030 +/- 0.006), (0.033 +/- 0.004), (0.033 +/- 0.007), (0.032 +/- 0.006), (0.044 +/- 0.012), (0.044 +/- 0.011), (0.042 +/- 0.007), (0.065 +/- 0.007), (0.044 +/- 0.014)h(-1) at duodenum, jejunum, ileum, respectively. The absorption rate of Rb1 was higher than R1 and Rg1.

CONCLUSIONAccording to the P and the logP, it can be conjectured that the absorption of R1 and Rg1 are better than Rb1. The absorption rate is decreased with the increase of the PNS concentration. Their absorption is the passive diffusion mechanism and other transport may also take part in the transport process. PNS is absorbed at all small-intestinal segments of rats, there are no significant differences between the three sections.

Animals ; Calibration ; Drug Stability ; Intestinal Absorption ; Intestine, Small ; metabolism ; Kinetics ; Male ; Oils ; chemistry ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Saponins ; chemistry ; metabolism ; Water ; chemistry

OBJECTIVETo study the effect of delayed sequential bone marrow transplantation on acute graft-versus-host disease (aGVHD) in major H-2 incompatible mouse transplantation.

METHODSC57BL/6 (H-2b) mice were used as donors and BALB/c (H-2d) mice as recipients. BALB/c mice were given 8.0 Gys total body irradiation (TBI) on day 0 and infused with a blend of bone marrow cells and spleen cells in different time. Transplantation was carried out as follows: group I TBI on day 0 and transplantation at 4 h after TBI; groups of II TBI on day 0 and transplantation at 4 h, d1, d2, d3 after TBI; groups III TBI on day 0 and transplantation at day 4 after TBI; groups IV TBI on day 0 and transplantation at day 4 through day 7 after TBI. Recipient's spleen H-2b cells were detected by flow cytometry and the level of serum cytokines (IL-2, IL4, IL-6, IL-10 and IFN-gamma) by ELISA. The survival, aGVHD and hematopoietic recovery were observed.

RESULTSaGVHD occurred in group I and the mice all died within 3 weeks after transplantation. The 60 day survival rates of groups of II and III were 30% and 50% respectively. The degree of aGVHD in group III was modest and the survival rate was higher than that in other groups (P <0.05). The peak time of IL-2, IFN-gamma, IL-4 and IL-10 in groups III and IV were later than that in group I. The levels of IL-4 and IL-10 in groups III and IV were higher than that in group I and for the levels of IL-2 and IFN-gamma were on the contrary (P < 0.05). The level of IL-6 in all groups peaked on day 5 to day 10 after TBI and was higher in group I than in others (P <0.05). In group IV the mean value of donor H-2b cells was (98.1 +/- 1.1)% on day 60 and WBC counts recovered normal on day 20.

CONCLUSIONSDelayed sequential transplantation can reduce the morbidity of aGVHD ,improve the survival rate and not affect the engraftment and reconstitution of hematopoiesis in mouse allo-BMT. The mechanism of aGVHD prevention may be related to the reducing of type 1 cytokines of T lymphocyte and the increasing of type 2 cytokines.

Animals ; Bone Marrow Transplantation ; immunology ; methods ; Disease Models, Animal ; Female ; Graft vs Host Disease ; immunology ; prevention & control ; Interleukin-2 ; blood ; Interleukin-4 ; blood ; Interleukin-6 ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL

OBJECTIVETo explore the silencing effect of short hairpin RNA (shRNA) on the CD28 of mice T lymphocytes by CD28-shRNA expressing plasmid evaluate the interfering effects (chronology and stability) mediated by shRNA and select out the most efficient CD28 shRNA sequence.

METHODSThree CD28 specific and one non-specific shRNA expressing plasmids were constructed and then transfected separately into mice spleen T lymphocytes. Non-transfected cells and non-specific shRNA were taken as controls. Inhibitory effect of CD28 shRNA was demonstrated by real-time quantitative PCR and Western blots. The sequence of the highest RNA interference (RNAi) efficacy was screened.

RESULTS(1) CD28 shRNA expressing plasmids were successfully constructed; (2) Three CD28 specific shRNAs effectively inhibited the expression of CD28 at the mRNA and protein levels, and there was a statistically significant difference comparing with the controls (P < 0.01): The copies of CD28 in mice spleen cells at the mRNA levels were persistently decreased by 99.62%, 99.89% and 99.80% respectively after 20 days, and so did at the protein level [(84.90 +/- 0.65)%, (96.49 +/- 0.03)%, (91.76 +/- 0.32)% respectively]. The highest inhibitory rate was in CD28 shRNA-2 group.

CONCLUSIONS(1) Specific shRNA can mediate long-term and stable silencing effects on CD28 gene; (2) shRNAs matching different sites of CD28 gene exert differential inhibitory effects.

Animals ; CD28 Antigens ; genetics ; Cells, Cultured ; Gene Expression Regulation ; Mice ; Mice, Inbred C57BL ; Plasmids ; genetics ; RNA Interference ; T-Lymphocytes ; metabolism ; Transfection

OBJECTIVETo explore the prophylaxis effect of pretreatment of allograft with methoxypolyethylene glycol-succinimidyl-propionic acid ester (mPEG-SPA) and anti-OX40L monoclonal antibody (McAb) on acute graft-versus-host disease (aGVHD) after allogeneic bone marrow transplantation (allo-BMT) in mice.

METHODSResponder splenocytes from C57BL/6 donor mice (H-2(b)) were co-cultured with stimulator splenocytes from BALB/c recipient mice (H-2(d)) for 7 days in the presence or absence of anti-OX40L McAb followed by mPEG-SPA chemical modification. Donor bone marrow cells plus the mixed culture of T-cells were then transplanted into lethally irradiated BALB/c mice. The BALB/c recipient mice were divided into four groups: group A (allo-BMT control group), group B(mPEG-SPA modification group), group C (anti-OX40L McAb pretreated group) and group D (mPEG-SPA and anti-OX40L McAb dual-treated group). Survival time and survival rate of the recipients were observed after allo-BMT. GVHD was assessed by clinical signs and histological changes of skin, liver and small intestines. Enzyme-linked immunosorbent assay (ELISA) was used to detect cytokines (IL-4, IL-10 and INF-gamma) production. Flow cytometry (FCM) analysis was used to detect allogeneic chimerism.

RESULTS(1) The mice in group A developed typical clinical signs of aGVHD and all mice died within 17 days after BMT with an average survival time (AST) of (12.1 +/- 5.5) days. The signs of aGVHD were less evident in mice of groups B, C and D, and their AST (36.2 +/- 24.9, 32.0 +/- 24.8 and 44.3 +/- 23.2 days, respectively) were all longer than that in group A (P < 0.05). AST of group D being the longest (P < 0.05). The survival rates at day 60 post-BMT in groups B, C and D were 50%, 41.7% and 66.7%, respectively. (2) Serum IFN-gamma level was increased after BMT in group A, and peaked in day 10 to day 15 post-BMT, while the level was decreased in groups B, C and D, reached the nadir on the day 10 post-BMT, with the lowest in group D (P < 0.01). After BMT, IL-4 and IL-10 levels were slightly decreased in group A, their levels were elevated in groups B and C (P < 0.05) and even more significantly increased in group D (P < 0.01). IL-4 and IL-10 levels peaked between day 10 and 15 post-BMT. (3) The average proportion of H-2(b) positive cells in recipient mice was 95% - 100% on day 60 post-BMT, with complete donor-type implantation.

CONCLUSIONCombination of mPEG-SPA and anti-OX40L McAb can block T-cell activated antigens and co-stimulatory pathway, regulate the T cells differentiation and induce the immune shift of Th0 cells toward Th2 cells. The immune tolerance induced by this method can significantly relieve aGVHD after allo-BMT.

Animals ; Bone Marrow Transplantation ; Graft vs Host Disease ; prevention & control ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transplantation, Homologous