Targeted delivery of RNAi to cancer cells using RNA-ligand displaying exosome

Exosome is an excellent vesicle for in vivo delivery of therapeutics, including RNAi and chemical drugs. The extremely high efficiency in cancer regression can partly be attributed to its fusion mechanism in delivering therapeutics to cytosol without endosome trapping. However, being composed of a lipid-bilayer membrane without specific recognition capacity for aimed-cells, the entry into nonspecific cells can lead to potential side-effects and toxicity. Applying engineering approaches for targeting-capacity to deliver therapeutics to specific cells is desirable. Techniques with chemical modification in vitro and genetic engineering in cells have been reported to decorate exosomes with targeting ligands. RNA nanoparticles have been used to harbor tumor-specific ligands displayed on exosome surface. The negative charge reduces nonspecific binding to vital cells with negatively charged lipid-membrane due to the electrostatic repulsion, thus lowering the side-effect and toxicity. In this review, we focus on the uniqueness of RNA nanoparticles for exosome surface display of chemical ligands, small peptides or RNA aptamers, for specific cancer targeting to deliver anticancer therapeutics, highlighting recent advances in targeted delivery of siRNA and miRNA that overcomes the previous RNAi delivery roadblocks. Proper understanding of exosome engineering with RNA nanotechnology promises efficient therapies for a wide range of cancer subtypes.

Puerarin protects against myocardial ischemia/reperfusion injury by suppressing NLRP3 inflammasome activation via TLR4/Myd88/NF-κB pathway in rats / 药学学报

This study was to investigate the protective effects of puerarin on myocardial ischemia/reperfusion (MI/R) injury and the underlying mechanism. The MI/R-model was established by ligating the left anterior descending artery (LAD) for 60 min followed by 24 h reperfusion, puerarin (10, 30, and 100 mg·kg-1) was orally administered 20 min before reperfusion. Cardiac function, myocardial infarct index, cardiac damage markers, inflammatory cytokines, and apoptosis index were measured to evaluate the protective effects of puerarin on MI/R injury. The activation of Nod-like receptor protein 3 (NLRP3) inflammasome and Toll like receptor 4 (TLR4)/myeloid differentiation factor 88 (Myd88)/nuclear factor kappa B (NF-κB) pathway were determined by Western blot. All animal experimental procedures were approved by the ethics committee of the Institute of Materia Medica, Peking Union Medical College, Chinese Academy of Medical Sciences. The results showed that puerarin could significantly improve cardiac function, reduce myocardial infarct size, decease the levels of lactic dehydrogenase (LDH), aspartate transaminase (AST), creatine kinase-MB (CK-MB), and cardiac troponin T (cTnT) and suppress cardiomyocyte apoptosis. Meanwhile, puerarin could notably decrease the levels of inflammatory cytokines such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α). Western blot analysis revealed that puerarin could downregulate the expression of TLR4, Myd88, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cleaved-caspase 1, cleaved-gasdermin-D (GSDMD), IL-1β, and IL-18, as well as the phosphorylation levels of inhibitor of NF-κB α (IκBα), IκB kinase β (IKKβ), and NF-κB. These findings demonstrated that puerarin could alleviate MI/R injury by suppressing NLRP3 inflammasome activation, possibly via TLR4/Myd88/NF-κB pathway.

Lack of association between mutation in IL36RN and palmoplantar pustular psoriasis in Chinese patients

Abstract Background: Palmoplantar pustulosis is considered to be a localized pustular psoriasis confined to the palms and soles. Mutation of the IL36RN gene, encoding interleukin-36 receptor antagonist (IL-36Ra), is associated with generalized pustular psoriasis, but IL36RN mutations in Chinese palmoplantar pustulosis patients have not previously been investigated. Objective: The aim of this study was to evaluate the mutation of IL36RN in Chinese patients with palmoplantar pustulosis. Methods: Fifty-one Han Chinese patients with palmoplantar pustulosis were recruited. All exons and exon-intron boundary sequences of IL36RN were amplified in polymerase chain reactions, and Sanger sequencing of the amplicons was performed. Results: Among the 51 palmoplantar pustulosis patients, four different single-base substitutions were identified in nine patients. The mutations were c.140A>G/p.Asn47Ser in five patients, c.258G>A/p.Met86IIe in two patients, and c.115+6T>C and c.169G>A/p.Val57IIe in one patient each. All mutations were heterozygous. Comparison with the human genome database and reported literature suggested that these variants may not be pathogenic mutations causing palmoplantar pustulosis. Furthermore, there was no difference in disease severity, onset age, or disease duration between patients with these heterozygous IL36RN variants and those without (p > 0.1). Study limitation: Lack of the further evaluation of IL36Ra protein in palmoplantar pustulosis lesions. Conclusions: The four variants of IL36RN identified did not appear to be associated with the specific phenotypes of palmoplantar pustulosis.

Total flavonoids of bugloss limits left ventricular remodeling after myocardial infarction in mice / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effects of total flavonoids of bugloss(TFB)on left ventricular (LV)remodeling after myocardial infarction(MI),LV size and function was compared in mice subjected to left anterior descending coronary artery ligation. METHODS 28 d after MI, the infarcted fraction of the LV and LV mass, systolic and diastolic function were measured. Capillary density and myocyte width in the nonischemic portion of the LV were also determined.RESULTS 28 d after MI,both groups had dilated LVs with decreased fractional shortening and lower ejection fractions. Although the infarcted size of the LV was similar in both groups,LV end-diastolic internal diameter,end-diastolic volume,and mass were lower, but fractional shortening, ejection fraction, and the maximum rate of developed LV pressure(dp/dtmax)were greater in TFB treated mice than in control mice.Impairment of diastolic func-tion, as measured by the time constant of isovolumic relaxation (t) and the maximum rate of LV pres-sure decay(dp/dtmin),was more marked in control mice than in TFB treated mice.Mortality after MI was greater in control mice than in TFB treated mice.In control mice,capillary density and myocyte width in the nonischemic portion of the LV did not differ before and 28 days after MI, whereas in TFB treated mice, capillary density increased and myocyte width declined after MI. CONCLUSION These results suggest that the presence of TFB limits LV dysfunction and remodeling in a murine model of MI in part by decreasing myocyte hypertrophy in the remote myocardium.

Distribution of Oral Pathogenic Bacteria and Analysis of Drug Resistance in the Infective Patients after Orthodontic Treatment in a Hospital from 2011 to 2016 / 中国药房

OBJECTIVE:To provide reference for clinical rational drug use and the prevention and treatment of drug-resistance bacteria.METHODS:A total of 148 patients with oral infection after orthodontic treatment were selected from a hospital during Jul.2011-Jul.2016.The distribution of pathogenic bacteria and drug resistance were analyzed retrospectively.RESULTS:Among 148 patients with oral infection,275 clinical specimens were detected,including 209 positive specimens with positive rate of 76.00％.A total of 332 pathogenic bacteria were detected,including 85 Gram-positive bacteria (25.60％) and 247 Gram-negative bacteria (74.40％).Top 7 isolated bacteria in the list of quantity were Actinobacillus pleuropneumoniae (54 strains,16.27％),Porphyromonas gingivalis (41 strains,12.35 ％),Tannerella forsythia (37 strains,11.14 ％),Streptococcus oralis (33 strains,9.94％),Klebsiella pneumoniae (30 strains,9.04％),Staphylococcus aureus (26 strains,7.83％) and Pseudomonas aeruginosa (25 strains,7.53％).Resistance rates of S.aureus to penicillin G,gentamicin,ciprofloxacin,oxacillin and tetracycline were all in high level (resistance rate＞50％),but it was sensitive to vancomycin and teicoplanin (resistance rate of 0).Enterococcus faecalis showed high resistance to penicillin G,erythromycin and oxacillin (resistant rate＞50％),but was sensitive to vancomycin and rifampicin (resistant rate of 0).K.Pneumoniae showed high resistance to gentamicin,ciprofloxacin,levofloxacin and cefazolin (resistant rate＞ 50％),but was sensitive to imipenem,ceftazidime,cefepime,ampicillin sodium and sulbactarn sodium,amoxicillin and clavulanate potassium (resistant rate＜ 10 ％).Resistant rates of P aeruginosa to gentamicin and levofloxacin were ≥ 80 ％,but it was sensitive to aztreonam (resistant rate of 8.00 ％).Resistant rate of Escherichia coli to piperacillin was 84.21％,but it was sensitive to imipenem and ampicillin sodium and sulbactam sodium (resistance rate of 5.26％).CONCLUSIONS:After orthodontic treatment,the pathogens of oral infection are various,mainly Gram-negative bacteria,and their drug resistance is not optimistic.The drugs with high sensitivity to the main pathogens include vancomycin,imipenem and enzyme inhibitor complex preparations,etc.Clinical attention should be paid to the cultivation of pathogenic bacteria and drag sensitivity test;according to the results of drug sensitivity test,targeted antibiotics should be selected to improve the antibacterial effect and delay the emergence of drug-resistant bacteria.

Study of sequential strategy for hepatitis B e antigen (HBeAg) positive chronic hepatitis B (CHB) patients with suboptimal early response to Peg-interferon-α / 中国基层医药

Objective To investigate the efficacy of entecavir (ETV) sequential therapy in the treatment of hepatitis B e antigen(HBeAg) positive chronic hepatitis B(CHB) patients with suboptimal early response to Peginterferon-α(Peg-IFN-α).Methods The cases of HBeAg-positive CHB who were treated with Peg-IFN-α for 12 to 24 weeks and serum HBsAg ＞ 20 000 IU/mL were enrolled into observation group.Treatment naive HBeAg positive CHB with serum HBsAg ＞ 20 000IU/mL were enrolled into control group.Both two groups received ETV for 96 weeks.Hepatitis B virus (HBV) virological and serological data were collected every 12 weeks.Results At the end of 48-week and 96-week,the rates of HBeAg seroconversion in the observation group were 23.3％ (10/43),30.2％ (13/43),respectively,which in the control group were 23.1％ (12/52),28.8％ (15/52),respectively.The HBsAg decline at 24-week was observed in both two groups.Conclusion Sequential strategy for patients with suboptimal early response to IFN is preferable.

Application of Ultrasonic Elastography in Diagnosis of Chronic Nonspecific Low Back Pain / 中国康复理论与实践

Objective To evaluate the value of ultrasonic elastography for diagnosis of chronic nonspecific low back pain. Methods From March to September, 2016, 32 patients diagnosed as chronic nonspecific low back pain and other 32 healthy people (controls) were measured lumbar erector spinae with ultrasonic elastography, and calculated Yang's modulus. The correlation between Yang's modulus and Visual Analogue Scale (VAS) or Japanese Orthopaedic Association (JOA) score in the patients were investigated with Pearson's analysis. Re-sults There was no significant difference in Yang's modulus between bilateral lumbar erector spinae in both the patients and the controls (t0.05), but it was more in the patients than in the controls (t=9.931, P<0.001). The Yang's modulus was positively correlated with VAS score in the patients (r=0.614, P<0.001), but not with the JOA score (r=-0.243, P=0.180). Conclusion Ultrasonic elastography can be applied to differentiate low back pain from the healthy, and measure the intensity of pain.

Efficacy and safety of Tripterygium wilfordii hook F versus acitretin in moderate to severe psoriasis vulgaris: a randomized clinical trial / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Few clinical trials have evaluated the efficacy and safety of Tripterygium wilfordii Hook F (TwHF) compared with acitretin in psoriasis. We aimed to compare the efficacy and safety of TwHF compared with acitretin in the treatment of moderate to severe psoriasis vulgaris.</p><p><b>METHODS</b>Adults with Psoriasis Area Severity Index (PASI) score ≥ 10 and psoriasis-affected body surface area ≥ 10% were randomized into a TwHF (20 mg, 3 times a day) or acitretin group (30 mg, once a day). The treatment course lasted for 8 weeks. Patients were assessed at baseline and at 2, 4, and 8 weeks. Laboratory tests were performed at baseline, week 4, and week 8. The data were analyzed using paired samples t-test or analysis of variance (ANOVA).</p><p><b>RESULTS</b>A total of 115 patients was enrolled (58 TwHF; 57 acitretin). The median PASI score improved in the TwHF group by 50.4% and in the acitretin group by 42.7%. There was no significant difference in median PASI improvement between two groups at 2, 4, and 8 weeks. There was also no significant difference in PASI 25, PASI 50, PASI 75, and PASI 90 response between the two groups at 2, 4, and 8 weeks. There was a significant increase in the level of aspartate transaminase and triglycerides in the TwHF group (P = 0.026 and P = 0.011, respectively). In the acitretin group, there was a significant increase in the level of alanine transaminase, cholesterol, and high-density lipoprotein (P = 0.030, P < 0.01, and P < 0.01, respectively).</p><p><b>CONCLUSIONS</b>There was no significant difference in treatment efficacy between the TwHF and acitretin groups within 8 weeks, but there were fewer treatment-related adverse events in the TwHF group.</p>

Serum levels of interleukin-36 and its receptor antagonist in patients with psoriasis and their correlations with disease severity / 中华皮肤科杂志

Objective To evaluate the relationship of serum levels of interleukin (IL)-36α,IL-36β and IL-36γas well as their receptor antagonist IL-36Ra with disease severity in patients with psoriasis.Methods Venous blood samples were collected from 45 patients with generalized pustular psoriasis (GPP),34 patients with psoriasis vulgaris (PV),and 37 healthy human controls.Enzyme-linked immunosorbent assay (ELISA) was performed to determine the serum levels of IL-36α,IL-36β,IL-36γ and IL-36Ra.Nonparametric Mann-Whitney test was conducted to compare the levels of IL-36 and IL-36Ra among these groups,and Spearman's rank correlation analysis to assess the relationship between the serum levels of IL-36 and IL-36Ra and disease severity.Results No statistical difference was observed in the serum levels of IL-36 or IL-36Ra among the patients with GPP,patients with PV,and healthy human controls.The serum levels of IL-36β and IL-36γ (given as the median ± interquartile range) were significantly higher in 27 patients with GPP during episodes of pustules ((12.101 ± 11.315) ng/L and (34.541 ± 15.580) ng/L respectively) and in 7 patients with severe GPP ((11.218 ± 9.318) ng/L and (38.536 ± 17.332) ng/L respectively) than in the healthy human controls ((5.355 ± 9.020) ng/L and (23.052 ± 22.410) ng/L respectively,P ＜ 0.05 or 0.01).The serum level of IL-36γwas positively correlated with that of IL-36β in patients with GPP,patients with PV,and the healthy human controls (r =0.85,0.86,0.91,respectively,all P ＜ 0.01),and both IL-36β and IL-36γserum levels were lowly and positively correlated with the severity of GPP (r =0.33,0.41,respectively,both P ＜ 0.05).A positive correlation was also observed between the serum level of IL-36β and psoriasis area and severity index (PASI) scores in patients with PV (r =0.54,P ＜ 0.01).Conclusions The serum levels of IL-363 and IL-36γ are lowly and positively correlated with disease severity in patients with GPP,suggesting that IL-36β and IL-36γplay an important role in the pathogenesis of GPP.

Effects of thalidomide on the expression and secretion of vascular endothelial growth factor and tumor necrosis factor-alpha by a human keratinocyte cell line HaCaT / 中华皮肤科杂志

Objective To investigate the effect of thalidomide on the proliferation of as well as the expression and secretion of vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α) by the human keratinocyte cell line HaCaT.Methods Cultured HaCaT cells were divided into several groups to be treated with dimethyl sulfoxide (negative control group) and various concentrations (0.01nmol/L-100 μmol/L) of thalidomide (experimental groups) respectively for 20 to 24 hours.Subsequently,water soluble tetrazolium-1(WST-1) assay was performed to estimate cellular proliferation,real time quantitative PCR to detect the mRNA expression of VEGF and TNF-α in HaCaT cells,and enzyme-linked immunosorbent assay (ELISA) to quantify the protein expressions of VEGF and TNF-α in the culture supernatants of HaCaT cells.Statistical analysis was done by one-way analysis of variance with least significant difference post hoc test.Results The survival rate of HaCaT cells was 74.3％,82.9％ and 90.8％ after 24-hour treatment with thalidomide of 100,10 and 1 μmol/L respectively,significantly lower than that in the negative control group (100％,all P ＜0.01).A significant decrease was induced in the mRNA expression (0.439-to 0.634-fold change,all P ＜0.01) and supematant level ((0.587-to 0.923-fold change,P ＜0.05) of VEGF in HaCaT cells by thalidomide of 0.01-100 nmol/L,as well as in the mRNA expression (0.493-to 0.587-fold change,P ＜0.05) and supernatant level (0.408-to 0.617-fold change,P ＜0.01) of TNF-α by thalidomide of 0.1-100 nmol/L.Conclusion Within a certain range of concentration,thalidomide could suppress the proliferation of,as well as the expression and secretion of VEGF and TNF-α by,HaCaT cells.

cpDNA non-coding sequence analysis of Pinellia ternata and its related species / 中草药

Objective: To provide the evidence for the identification and genetic diversity of Pinellia ternata by studying cpDNA non-coding sequences of P. ternata and its related species. Methods: Besides P. cordate and P. pedatisecta, 43 P. ternata samples were collected from the main habitats in China. psbK-psbI and atpF-atpH sequences in leaf genome DNA were cloned by PCR. Comparative analysis was carried out by bioinformatics software. Results: The sequence length of atpF-atpH in P. ternata was 337-342 bp and conservative. The numbers of variable sites and parsimony information sites were only 7 and 1, respectively. The genetic distance was 0-0.024.The length of psbK-psbI was 432-435 bp, with 37 variable sites, including 17 information sites, and the genetic distance was 0-0.069.Cluster analysis was not consistent with the phenotype or the geographical distributions. Conclusion: The discrimination of psbK-psbI is better than that of atpF-atpH, and there are more mutation sites in psbK-psbI sequence among species in P. ternata.

Neuroprotective effects and mechanisms of Chuanxiong Chatiao pulvis against MPTP-induced dopaminergic neurotoxicity in mice model of Parkinson's disease / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the neuroprotective effects and mechanisms of Chunxiong Chatiao pulvis (CXCT) on the MPTP-induced dopaminergic neurodegneration in mice model of PD.</p><p><b>METHOD</b>Three to five months old male mice were randomly divided into following six treatment groups (NS + NS, 50 mg x kg(-1) CXCT + NS, 25 mg x kg(-1) CXCT + MPTP, 50 mg x kg(-1) CXCT + MPTP, 75 mg x kg(-1) CXCT + MPTP, NS + MPTP) (n = 8/group). For behavioral measurement, the mice were evaluated for their motor coordination by pole-test; Striatal dopamine content of mice was measured using standard reverse-phase HPLC; Number of tyrosine hydroxylase (TH)-positive cells in the substantia nigra of mice was detected by immunohistochemistry; SOD activity in the substantia nigra of mice were measured by fluorimetry.</p><p><b>RESULT</b>The score of pole test (25, 50, 75 mg x kg(-1)), the striatal dopamine contents (75 mg x kg-1), the residual DA contents (75 mg x kg(-1)) and the SOD activity in substantia nigra (50, 75 mg x kg(-1)) all showed that, the CXCT-pretreated groups significantly higher than that of the group NS + MPTP (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>Chuanxiong Chatiao pulvis improved the motor deficit in MPTP-induced mice, and attenuate MPTP-induced dopaminergic neurodegeneration in mice. Importantly, the neuroprotective effect of Chuanxiong Chatiao pulvis against dopaminergic neurodegeneration may be associated with its strong antioxidant capacity in vivo.</p>

Oxytocin inhibits peripheral stimulation-induced LTP and FOS protein expression in hippocampus of rats / 基础医学与临床

Objective To investigate the effects of Oxytocin(OT) on hippocampal long-term potentiation(LTP) and FOS protein expression induced by peripheral stimulation.Methods Single stimulation pulses were delivered to the left sciatic nerves to evoke the field excitatory postsynaptic potentials (fEPSPs) in the right hippocampal CA1. Tetanic stimulation was used to induce hippocampal LTP. Different groups of rat were given NS,OT,Oxytocin antagonist-Atosiban + OT before tetanic stimulation,into lateral ventricle(LV) respectively. Expression of FOS protein was compared among the groups by histopathological and immunohistochemistry. Results Single stimulation evoked fEPSPs in hippocampal CA1,which average latency was (171.9?33.1)ms and average amplitude was (25.7?8.4)?V. Tetanic stimulation induced hippocampal LTP and increased the expression of FOS protein. Intracerebroventricular injection of OT inhibited hippocampal LTP and decreased the expression of FOS protein. This effect of OT was blocked by the pretreatment with Atosiban. Conclusion The results suggested that OT may play an inhibitory role in leaning and memory of rats and the effect is mediated by OT receptor.

Long-term effect of lamivudine combined with interferon alpha treatment for chronic hepatitis B virus infection / 中国实用内科杂志

Objective To evaluate the efficacy of 2-year treatment of interferon alpha combined with lamivudine therapy in chronic hepatitis B patients. Methods From 1999 to 2001 87 patients were enrolled and randomized into two groups.36 patients received interferon alpha and lamivudine simultaneously for 26 weeks,then lamivudine alone for 76 weeks.51 patients received lamivudine alone for 104 weeks.The efficacy was evaluated by biochemical and virological parameters.The YMDD motif was detected in both groups. Results There was no difference between two groups in HBV DNA response ( P =0.24).Proportion of HBeAg/Anti-HBe seroconversion in combination group was significantly higher than lamivudine group (38.9% vs 17.6%)( P =0.03).HBV YMDD mutation rate in combination group was lower than in lamivudine group (22.2% vs 43.1%, P =0.04). Conclusion The efficacy of combination therapy appeares to be better than lamivudine monotherapy.Combination treatment may delay or diminish the development of HBV YMDD mutation.

Interferon-? combined with lamivudine in the treatment of chronic hepatitis / 中华传染病杂志

1.0 ?10 7 copies/ml than those with lower HBV DNA levels both in combination group and lamivudine group (52.0% vs 25.8%, P= 0.04; 63.2% vs 28.6%, P= 0.03). Necro-inflammatory activity, fibrosis and the expression of HBV DNA showed no correlation with response rate in combination group. Among those who had higher levels of HBV DNA expression, seroconversion rate in combination group is higher than that in lamivudine group (46.2% vs 9.5%, P= 0.03). Conclusions The efficacy of combination therapy appeared to be better in patients with at least moderately elevated baseline ALT, high level of HBV DNA as well as HBV DNA expression in hepatocytes. Combination therapy may delay or diminish the development of YMDD mutation-related resistance to lamivudine.