Understanding molecular characteristics of extracellular vesicles derived from different types of mesenchymal stem cells for therapeutic translation.

Mesenchymal stem cells (MSCs) have been studied for decades as candidates for cellular therapy, and their secretome, including secreted extracellular vesicles (EVs), has been identified to contribute significantly to regenerative and reparative functions. Emerging evidence has suggested that MSC-EVs alone, could be used as therapeutics that emulate the biological function of MSCs. However, just as with MSCs, MSC-EVs have been shown to vary in composition, depending on the tissue source of the MSCs as well as the protocols employed in culturing the MSCs and obtaining the EVs. Therefore, the importance of careful choice of cell sources and culture environments is receiving increasing attention. Many factors contribute to the therapeutic potential of MSC-EVs, including the source tissue, isolation technique, and culturing conditions. This review illustrates the molecular landscape of EVs derived from different types of MSC cells along with culture strategies. A thorough analysis of publicly available omic datasets was performed to advance the precision understanding of MSC-EVs with unique tissue source-dependent molecular characteristics. The tissue-specific protein and miRNA-driven Reactome ontology analysis was used to reveal distinct patterns of top Reactome ontology pathways across adipose, bone marrow, and umbilical MSC-EVs. Moreover, a meta-analysis assisted by an AI technique was used to analyze the published literature, providing insights into the therapeutic translation of MSC-EVs based on their source tissues.

FAP is critical for ovarian cancer cell survival by sustaining NF-κB activation through recruitment of PRKDC in lipid rafts.

Fibroblast activation protein (FAP) is tumor-specific and plays an important role in tumorigenecity. However, agents against its enzymatic activity or extracellular presence were unsuccessful in the clinic for undefined reasons. Here we show that FAP expression is higher in advanced ovarian cancer and is only detected in invasive ovarian cancer cells. Silencing FAP induces apoptosis and FAP's enzymatic activity is dispensable for cell survival. To elucidate the cause of apoptosis, we find that NF-κB activity is diminished when FAP is depleted and BIRC5 (survivin) acts downstream of FAP-NF-κB axis to promote cell survival. To uncover the link between FAP and NF-κB activation, we reveal that PRKDC (DNA-PK, DNA-dependent protein kinase) forms complex with FAP and is required for NF-κB activation and cell survival. Remarkably, FAP-PRKDC interaction occurs only in lipid rafts, and depleting FAP prevents lipid raft localization of PRKDC. Given the known ability of PRKDC to direct NF-κB activation, these results suggest that FAP recruits PRKDC in lipid rafts for NF-κB activation. FAP's non-enzymatic role and functioning from lipid rafts for cell survival also offer an explanation on the failure of past FAP-targeted therapies. Finally, we demonstrate that EpCAM aptamer-delivered FAP siRNA impeded intraperitoneal xenograft development of ovary tumors.

CRISPR-Cas9 Engineered Extracellular Vesicles for the Treatment of Dominant Progressive Hearing Loss.

Clinical translation of gene therapy has been challenging, due to limitations in current delivery vehicles such as traditional viral vectors. Herein, we report the use of gRNA:Cas9 ribonucleoprotein (RNP) complexes engineered extracellular vesicles (EVs) for in vivo gene therapy. By leveraging a novel high-throughput microfluidic droplet-based electroporation system (µDES), we achieved 10-fold enhancement of loading efficiency and more than 1000-fold increase in processing throughput on loading RNP complexes into EVs (RNP-EVs), compared with conventional bulk electroporation. The flow-through droplets serve as enormous bioreactors for offering millisecond pulsed, low-voltage electroporation in a continuous-flow and scalable manner, which minimizes the Joule heating influence and surface alteration to retain natural EV stability and integrity. In the Shaker-1 mouse model of dominant progressive hearing loss, we demonstrated the effective delivery of RNP-EVs into inner ear hair cells, with a clear reduction of Myo7ash1 mRNA expression compared to RNP-loaded lipid-like nanoparticles (RNP-LNPs), leading to significant hearing recovery measured by auditory brainstem responses (ABR).

Stimulation of mGluR5 by VU0360172 protected against germinal matrix hemorrhage in neonatal rats / 中国药理学通报

Aim To investigate the protective effect of mGluR5 activated by VU0360172 on germinal matrix hemorrhage in neonatal rats.Methods Seven day- old SD rats were randomly divided into Sham, GMH, and low-, medium-, and high-dose groups.The model was established by intracerebral injection of collagenase W-S.Then three doses of VU0360172 were injected intraperitoneal

Prognostic role of cyclin D2/D3 in multiple human malignant neoplasms: A systematic review and meta-analysis.

Cyclin D2/D3 (CCND2/3) are core components of the machinery that drives cell cycle progression and therefore, are associated with tumorigenesis. Currently, there are contradictory evidences on the function of CCND2/3 in tumorigenesis. Thus, we conducted a comprehensive meta-analysis to derive a precise predictive value of CCND2/3 in various tumors. We searched PubMed, EMBASE, Web of Science for eligible studies up to October 8, 2018. Pooled hazard ratios (HRs) with 95% confidence intervals (CIs) of OS or DFS/PFS/RFS were calculated using Forest plot analysis to demonstrate their associations. A total of 14 studies were ultimately included in this meta-analysis. Our results indicated CCND2/3 played an oncogenic role in all of the cancer patients (CCND2: pooled HR = 2.21, 95% CI: 1.67-2.93; CCND3: pooled HR = 2.29, 95% CI: 1.05-5.03). In tumor subgroup, CCND2 was associated with shorter OS in patients with gastric cancer (HR = 2.20, 95% CI: 1.66-2.92), whereas it might be a tumor suppressor in NSCLC (HR = 0.28, 95% CI: 0.12-0.64). In addition, CCND3 was correlated to reduced OS in breast cancer patients (HR = 1.64, 95% CI: 1.07-2.52) and shorter DFS/PFS/RFS in bladder cancer patients (HR = 4.60, 95% CI: 1.89-12.57). Taken together, CCND2/3 could be the promising biomarkers for predicting the prognosis of patients with malignant neoplasms.

A Novel Inhibitor of Homodimerization Targeting MyD88 Ameliorates Renal Interstitial Fibrosis by Counteracting TGF-ß1-Induced EMT in Vivo and in Vitro.

BACKGROUND/AIMS: The TLR/MyD88/NF-κB signaling pathway has been successfully used to treat renal interstitial fibrosis (RIF). However, the exact therapeutic mechanism is still unknown. Here, we assessed the therapeutic efficacy of TJ-M2010-2, a small molecular compound that inhibits MyD88 homodimerization, in RIF induced by ischemia reperfusion injury (IRI). METHODS: In vivo, RIF was induced in mice by IRI, and the mice were prophylactically treated with TJ-M2010-2. In vitro, HK-2 cells were incubated with TGF-ß1 to induce EMT, and the cells were pretreated with TJ-M2010-2. RESULTS: We found that, compared with the IRI group, the TJ-M2010-2 group showed marked attenuation of RIF and renal function injury; decreased expression of TGF-ß1, α-SMA, vimentin, MMP2 and MMP9; and increased E-cadherin expression. Furthermore, TGF-ß1-induced EMT was blocked by TJ-M2010-2 in HK-2 cells, as evidenced by blocked morphologic transformation, restored E-cadherin expression and inhibited α-SMA expression. In addition, compared to the TGF-ß1 group, the TJ-M2010-2 group showed profound inhibition of the expression of TRAF6, p65 and Snail and upregulation of the expression of IκBα. CONCLUSION: This MyD88 inhibitor may be a potential therapeutic agent to ameliorate RIF.

The feasibility of 1-stop examination of coronary CT angiography and abdominal enhanced CT.

This study aims to evaluate the feasibility of performing coronary computed tomography angiography (CCTA) and abdominal enhanced computed tomography (CT) with 1-time injection of the agent.CCTA images (right coronary artery, left anterior descending coronary artery, and left circumflex coronary artery) were collected from 20 patients who completed a 1-stop combined examination of CCTA and abdominal enhanced CT (group A), 20 patients who only underwent abdominal enhanced CT (group B1), and 20 patients who only underwent CCTA (group B2). These images were interpreted using the 5-point Likert scale system by 2 experienced radiologists, and abdominal images were observed for breathing artifact. CT value, signal-to-noise ratio (SNR), and CTDI were recorded and compare among the 3 groups.The difference in image quality of the coronary and total volume of the contrast agent between group A and group B1 was not statistical significant (P > .05). The CT value and SNR in group B1 (CCTA) (CT: 394.65 ±â€Š59.23, SNR: 17.38 ±â€Š4.13) increased, compare with Group A (CT: 360.35 ±â€Š34.16, SNR: 13.76 ±â€Š1.84, P = .03, .01), while CTDI was undifferentiated between group A (17.14 ±â€Š6.20) and group B1 (18.38 ±â€Š9.79) (P = .64). The difference in CT value and SNR at the arterial phase and CT value at the venous phase between group A (abdomen) and group B2 were statistically significant, the CTDI in group A (9.09 ±â€Š1.05) increased, compared with group B2 (8.23 ±â€Š1.33) (P = .03), and SNR at the venous phase in group B2 (12.50 ±â€Š2.43) increased, compared with group A (10.89 ±â€Š2.03) (P = .03).Revolution CT can capture full images and very rapidly switch to the scan mode, enabling a 1-stop axial CCTA and enhanced helical abdominal scan. The 1-stop combined scan resulted in a satisfactory image quality, which reduced the contrast agent dose and simplified the workflow.The 1-stop combined scan allows for the high success rate of the examination, reduces the number of examinations, and decreases the dose and risk of injection of the contrast agent. This would be helpful for patients to obtain diagnostic images in time.

Formula Optimization of Captopril Timing Osmotic Pump Tablets / 中国药房

OBJECTIVE:To optimize the formula of Captopril timing osmotic pump tablets. METHODS:Using accumulative release rate (Q) as index, single factor test was used to investigate the effects of blocking layer coating weight gain, semipermeable membrane coating weight gain,the type of polyepoxide (PEO),the amount of PEO (3 × 105) and HPMC in drug bearing layer,the amount of PEO (7 × 106) and NaCl in booster layer on drug release of Captopril timing osmotic pump tablets. Based on single factor investigation,using comprehensive score of release behavior(L)as index,the amount of PEO(3×105)and HPMC in drug bearing layer,the amount of PEO(7×106)and NaCl in booster layer as factors,L9(43)orthogonal test was used to optimize the formula of tablet core validation test was conducted. RESULTS:The optimal formula of tablet core included PEO(3× 105)71 mg and HPMC 15 mg in drug bearing layer,PEO(7×106)61 mg and NaCl 18 mg in booster layer,coating weight gain 7％ and semipermeable membrane coating weight gain 10％ in blocking layer. The osmotic pump tablet prepared by optimized formula released after 4 h;in vitro drug release regression equation was Q=5.118t-21.441(R2=0.9956),which was in line with zero-order release characteristics. CONCLUSIONS:The optimal formula is stable,feasible and controllable in quality,and can provide reference for further development of Captopril timing osmotic pump tablets.

Short-term MyD88 inhibition ameliorates cardiac graft rejection and promotes donor-specific hyporesponsiveness of skin grafts in mice.

Recognition of evolutionarily conserved ligands by Toll-like receptors (TLRs) triggers signaling cascades in innate immune cells to amplify adaptive immune responses. Nearly all TLRs require MyD88 to transduce downstream signaling. MyD88 deficiency has been shown to promote the allograft acceptance in mice. However, direct evidence for therapeutic potential of MyD88 inhibitors remains lacking. Herein, we used a MyD88 inhibitor, namely ST2825, to explore its therapeutic potential and mechanisms in fully allogeneic skin and heart transplant models. Phenotypic maturation of dendritic cells stimulated by TLR ligands was alleviated by ST2825 in parallel with reduced T-cell proliferation in vitro. A short-course treatment with ST2825 significantly prolonged cardiac graft survival (mean survival time = 18.5 ± 0.92 days vs. 7.25 ± 0.46 days). ST2825-treated group had significantly reduced proinflammatory cytokines in allografts compared with control group. ST2825 combined with anti-CD154 induced long-term skin allograft acceptance in about one-third of recipients (>100 days). 'Skin-tolerant' recipients showed attenuated donor-specific IFN-γ responses, intact IL-4 responses, and compromised alloantibody responses. We conclude that MyD88 inhibitor ST2825 attenuates acute cardiac rejection and promotes donor-specific hyporesponsiveness in stringent skin transplant models. The direct evidence suggests that pharmacological inhibition of MyD88 hold promising potential for transplant rejection.

Survey of Clinical Pharmacy Work Status in Medical Institutions of Guizhou Province / 中国药师

Objective:To know the work status of clinical pharmacy in medical institutions of Guizhou province. Methods:Ques-tionnaires were used to analyze the situation of clinical pharmacy in 108 medical institutions of Guizhou province. Results: A total of 246 questionnaires were taken back, and among the 231 valid questionnaires were received including gradeⅡor above hospitals. The main contents of clinical pharmacy work carried out in medical institutions included 7 aspects: pharmacists ’ participation in ward rounds, which accounted for 47. 11%; pharmacists’ participation in case consultation, which accounted for 16. 65%; pharmacists’ participation in teaching practice, which accounted for 38. 84%; pharmacists’ participation in prescription evaluation and analysis, which accounted for 72. 73%;pharmacists’ participation in antimicrobial drug monitoring and drug use evaluation, which accounted for 62. 37%;pharmacists’ participation in drug counsultation and education, which accounted for 58. 68%;pharmacists’ participation in adverse drug reaction monitoring and supervision, which accounted for 77. 32%. Conclusion:The development of clinical pharmacy in Guizhou province still lags behind, and the number of clinical pharmacists is insufficient, which can’ t meet the growing demand for personalized medicine. In particular, the development of clinical pharmacy is restricted by the limited pharmaceutical service. The cog-nition degree of pharmacist group in Guizhou province has been improved. However, the number and the service quality of clinical pharmacists need to be improved further.

Rice grassy stunt virus nonstructural protein p5 serves as a viral suppressor of RNA silencing and interacts with nonstructural protein p3.

Rice grassy stunt virus (RGSV), a member of the genus Tenuivirus, causes serious rice disease in Southeast Asian countries. In this study, a green fluorescent protein (GFP)-based transient expression assay was conducted to show that p5, encoded on RNA5 in the viral sense, is a viral suppressor of RNA silencing (VSR). Protein-protein interactions (PPIs) between p5 and all RGSV proteins except pC1 and pC2 were investigated using Gal4-based yeast two-hybrid (Y2H) experiments. The results demonstrated that p5 interacts with itself and with p3 encoded on RNA3 in the viral sense. p5-p5 and p5-p3 interactions were detected by bimolecular fluorescence complementation (BiFC) assay, and the p5-p3 interaction was confirmed by subcellular co-localization and co-immunoprecipitation (Co-IP) assays. Using the Y2H system, we demonstrated that the p5-p3 interaction requires both the N-terminal (amino acid residues 1 to 99) and C-terminal (amino acid residues 94 to 191) domains of p5. In addition, either p5 or p3 could enhance the pathogenicity of potato virus X (PVX) in Nicotiana benthamiana plants. A much more significant enhancement of PVX pathogenicity and accumulation was observed when p5 and p3 were expressed together. Our data also showed that RGSV p3 does not function as a VSR, and it had no effect on the VSR activity of p5 or the subcellular localization pattern of p5 in plant cells from Nicotiana benthamiana.

Effective evaluation of presurgical nasoalveolar molding devices in the treatment of complete unilateral cleft lip and palate patients / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the orthopedic effect of presurgical nasoalveolar molding (PNAM) devices on the palatal deformities in unilateral complete cleft lip and palate (UCCLP) patients.</p><p><b>METHODS</b>Three groups with 19 patients each were studied. All samples in groups A and B were non-syndromic UCCLP children. Group A was treated with PNAM prior to operation. Group B was untreated prior to operation. Samples in group C were normally developed nose and lip palate infants aged three months. The orthotopic palate photos before and after PNAM treatment for group A, as well as pre-operative photos of groups B and group C, were taken and measured. All statistics were analyzed using SPSS 21.0.</p><p><b>RESULTS</b>PNAM treatment significantly increased the AW, AC, and PA of UCCLP patients (P < 0.05), whereas CPW, CWA, CWAS, CWAH, PMD, and CA significantly decreased (P < 0.05). However, no significant difference was observed with the cases in group C (P < 0.05). The AW, CPW, CA, and PA of the patients in group B significantly increased compared with the cases in group A before PNAM treatment (P < 0.05). Multivariate analysis of variance indicated that TW had no statistically significant difference among the three groups (P > 0.05).</p><p><b>CONCLUSION</b>PNAM treatment is a non-surgical early treatment for the effective improvement of palatal primary deformities in UCCLP patients.</p>

Theoretical study on chiral recognition mechanism of methyl mandelate enantiomers on permethylated ß-cyclodextrin.

Host-guest interactions of permethylated ß-cyclodextrin (PM-ß-CD) with methyl mandelate enantiomers ((R/S)-MMA) were simulated using semiempirical PM3 and ONIOM (B3LYP/6-31G(d):PM3) method. The chiral recognition mechanism of (R/S)-MMA enantiomers on PM-ß-CD was investigated. The binding energies for all orientations considered in this research are reported. The most stable geometry structures of the two complexes are different. The benzene ring of (R)-MMA locates horizontally approximately on the wider edge of the PM-ß-CD cavity, but the aromatic ring of (S)-MMA is deeply included into the hydrophobic cavity. Furthermore, the results of NBO analysis show that the main driving forces in the inclusion process of PM-ß-CD with (R/S)-MMA are hydrogen bonding interaction, dipole-dipole interaction, charge-transfer and hydrophobic interaction. The stabilization energy of the (R)-MMA/PM-ß-CD complex is lower than that of the (S)-MMA/PM-ß-CD complex. Moreover, the chiral carbon in MMA of (R/S)-MMA/PM-ß-CD complexes are close to the C2 and C3 in the glucose unit. The chiral recognition mechanism is thus closely related to the chiral environment provided by C2 and C3 in the glucose unit and the degree of (R/S)-MMA and PM-ß-CD inclusion.

[Simultaneous determination of five lignans in Schisandra chinensis by HPLC].

OBJECTIVE: To develop a HPLC quantitative method for determination of schisandrin, schisandrol B, schisantherin A, deoxyschisandrin and gamma-schisandrin in Fructus Schisandrae. METHOD: A symmetry C18 column (4.6 mm x 250 mm, 5 microm) was used with methanol (A) and water (B) as mobile phases, in gradient elution. The gradient program was as follows: 0-36 min, changed from 60% A to 66% A, 36-65 min, to 80% A, 65-70 min, kept for 80% A, 70-75 min, to 100% A. The flow rate was 0.5 mL x min(-1) and detection wavelength was set at 254 nm. RESULT: The linearities of schisandrin, schisandrol B, schisantherin A, deoxyschisandrin and gamma-schisandrin were in the ranges of 0.0214. 160 (r = 0.9999), 0.020-4.000 (r = 0.9999), 0.021-4.240 (r = 0.9999), 0.020-3.960 (r = 0.9999) and 0.021-4.200 (r = 0.9999). The average recoveries were 104.8%, 104.2%, 102.7%, 104.6%, 104.5%, respectively. CONCLUSION: The method developed in this study was reliable, and can be used for the quality control of the fruits of S. chinensis.