Objective To investigate the influence of surface grafting density of polyethylene glycol (PEG) molecules on the interactions of nanoparticle with the biomembrane.Methods The dynamics of polymer as well as its shielding effect on the wrap of the membrane were investigated using coarse-grained (CG) molecular dynamics simulations.A coarse grained model force field,MARTINI,was employed for its ability to reproduce experimental properties of various lipid dynamics and some kinds of polymer as well.Results As a result,structural variations and energy changes of grafted PEG molecules on the nanoparticles,adsorbing dynamics of PEGs on the membrane as well as the wrap rates of the membrane were obtained to shed light on the PEGylation-mediated shielding mechanism in the adsorption of PEGylated nanoparticles on the membrane.Conclusions The surface grafting density of PEGylated nanoparicles has been found to play a crucial role in the stealth shielding behavior of PEGylated nanocarriers.These studies are consistent with experimental observations and to some extent give a molecular level interpretation of the macroscale observations of prolonged circulation half-life of stealth nanocarriers.

OBJECTIVE:To provide reference for revising the standards of primaquine phosphate in International Pharmacopoe-ia (the fifth edition). METHODS:Based on inspection standards for primaquine phosphate and its preparations in different coun-tries,HPLC,perchloric acid potentiometric titration and sodium nitrite dead-stop titration were adopted to determine and compare the contents of primaquine phosphate. RESULTS:In HPLC,the linear range of primaquine phosphate was 128.1-384.2 μg/ml(r=0.999 9);the limit of detection was 0.18 μg/ml,the limit of quantification was 0.59 μg/ml;RSDs of precision,stability and repro-ducibility tests were lower than 1%;recovery was 99.42%-101.14%(RSD=0.6,n=9);destructive test and durability test showed good specificity;the contents of 3 batches of samples were 92.1%,92.3% and 92.0%,respectively. In perchloric acid potentiomet-ric titration,RSDs of precision and reproducibility tests were 1.6%;the titration jump was not obvious;the contents were 99.1%, 99.7% and 98.7%,respectively. In sodium nitrite dead-stop titration,RSDs of precision and reproducibility tests were lower than 1%;the titration jump was obvious;the contents were 96.9%,97.1% and 96.7%,respectively. Sodium nitrite external indicator ti-tration showed better precision than dead-stop titration;the contents were 99.6%,100.0% and 99.5%,respectively. CONCLU-SIONS:Perchloric acid potertiometric titration is simple and rapid,but with relatively poor precision and reproducibility,and it contains quinocide. Sodium nitrite dead-stop titration is simple with good precision and reproducibility but long reaction time,and it also contains quinocide,the HPLC method shows good separation and high accuracy,can effectively separate the quinocide in primaquine phosphate,and it is suitable for the quality control of primaquine phosphate.

ObjectiveTo investigate the relationship between the clinical type of attack and electroencephalogram (EEG) features of cerebral palsy (CP) children with epilepsy (EP).MethodsThe EEGs of 36 CP cases with EP were recorded under natural circumstances.Results20 cases (55.6%) had focal high amplitude spikes and wave complexes; 4 cases (11.1%) had typical hypsarrhythmia; 3 cases (8.3%) had varied hypsarrhythmia. Among them, the spikes and wave complexes of 9 cases were bilateral.ConclusionThe most of the clinical attack and EEG characters of CP children with EP are partial type.

OBJECTIVE: To evaluate and compare whole exome sequencing (WES) and targeted panel sequencing in the clinical molecular diagnosis of the Chinese families affected with inherited retinal dystrophies (IRDs). METHODS: The clinical information of 182 probands affected with IRDs was collected, including their family history and the ophthalmic examination results. Blood samples of all probands and their relatives were collected and genomic DNA was extracted by standard protocols. The first 91 cases were subjected to the WES and the other 91 cases were subjected to a specific hereditary eye disease enrichment panel (HEDEP) designed by us. All likely pathogenic and pathogenic variants in the candidate genes were determined by Sanger sequencing and co-segregation analyses were performed in available family members. Copy number variations (CNVs) detected by HEDEP were further validated by multiplex ligation-dependent probe amplification (MLPA). As PRGR ORF15 was difficult to capture by next generation sequencing (NGS), all the samples were subjected to Sanger sequencing for this region. All sequence changes identified by NGS were classified according to the American College of Medical Gene-tics and Genomics and the Association for Molecular Pathology (ACMG/AMP) variant interpretation guidelines. In this study, only variants identified as pathogenic or likely pathogenic were included, while those variants of uncertain significance, likely benign or benign were not included. RESULTS: In 91 cases with WES, pathogenic or likely pathogenic variants were determined in 30 cases, obtaining a detection rate of 33.00% (30/91); While in 91 cases with HEDEP sequencing, pathogenic or likely pathogenic variants were determined in 51 cases, achieving the diagnostic rate of 56.04% (51/91), and totally, the diagnostic rate was 44.51%. HEDEP had better sequencing coverage and read depth than WES, therefore HEDEP had higher detection rate. In addition, HEDEP could detect CNVs. In this study, we detected disease-causing variants in 29 distinct IRD-associated genes, USH2A, ABCA4 and RPGR were the three most common disease-causing genes, and the frequency of these genes in Chinese IRDs population was 11.54% (21/182), 6.59% (12/182) and 3.85% (7/182), respectively. We found 43 novel variants and 6 cases carried variants in RPGR ORF15. CONCLUSION NGS in conjunction with Sanger sequencing offers a reliable and effective approach for the genetic diagnosis of IRDs, and after evaluating the pros and cons of the two sequencing methods, we conclude that HEDEP should be used as a first-tier test for IRDs patients, WES can be used as a supplementary molecular diagnostic method due to its merit of detecting novel IRD-associated genes if HEDEP or other methods could not detect disease-causing va-riants in reported genes. In addition, our results enriched the mutational spectra of IRDs genes, and our methods paves the way of genetic counselling, family planning and up-coming gene-based therapies for these families.
DNA Copy Number Variations ; Humans ; Mutation ; Pedigree ; Retinal Dystrophies/genetics* ; Whole Exome Sequencing

Objective To investigate the inhibitory effects and mechanism of ginsenoside Rg3 on vasculogenic mimicry of human breast cancer MCF-7 cell line. Methods The MCF-7 cells at logarithmic growth phase were obtained, and were cultured with different concentrations (0, 20, 50, 100, 150 and 300 mg/L) of ginsenoside Rg3. Cells cultured without Rg3 were served as controls. The IC50 were determined by CCK8 assay and anti-angiogenic effects were performed for testing the potential of tube-like structure (TLSs) formation. The expression levels of VEGF-A, MMP 9 and HIF-1αwere detected by Western blotting and real-time PCR. The secreted contents of VEGF-A and MMP9 were detected by enzyme linked immunosorbent assay (ELISA). Results The ginsenoside Rg3 suppressed the proliferation of MCF-7 cells in a dose-dependent manner, in which IC50 was (115.34±8.50) mg/L. The formation numbers of TLSs in MCF-7 cells were significantly inhibited by Rg3 in concentration dependent manner in 50 mg/L, 100 mg/L and 150 mg/L for (19.0 ± 1.0), (15.0 ± 1.5), and (10.0±1.7) vs. controls (22.0±1.8, F=150.805, P<0.05). The mRNA and protein expression levels of VEGF-A, MMP9 and HIF-1αprotein were inhibited by 50 mg/L,100 mg/L and 150 mg/L Rg3 vs. controls (P<0.05). Meanwhile, the contents of VEGF-A in MCF-7 cell supernatant was down-regulated by 50 mg/L,100 mg/L and 150 mg/L Rg3 vs. controls (P<0.05). The contents of MMP-9 in MCF-7 cell supernatant was down-regulated by 100 mg/L and 150 mg/L Rg3 vs. controls (P<0.05). There was no significant difference in MMP-9 expression between 50 mg/L group and control group. Conclusion The ginsenoside Rg3 is able to inhibit the vasculogenic mimicry of MCF-7 cells, which may be related with the down-
regulation of VEGF-A, MMP9 and HIF-1α.

OBJECTIVE:To establish a method for the main active components in Citrus chacheiensis with different storage time (1-19 years). METHODS:HPLC was conducted to determine the content of hesperidin:the column was Diamonsil C18 with mobile phase of methanol-acetic acid-water(35∶4∶61,V/V/V)at a flow rate of 1.0 ml/min,detection wavelength was 283 nm,col-umn temperature was 25 ℃,and the injection volume was 5 μl;the contents of nobiletin and tangeretin:the column was Diamon-sil C18 with mobile phase of water-methanol(gradient elution)at a flow rate of 1.0 ml/min,detection wavelength was 326 nm,col-umn temperature was 25 ℃,and the injection volume was 10 μl;and the content of synephrine:the column was Diamonsil C18 with mobile phase of methanol-potassium dihydrogen phosphate solution(taking 0.6 g potassium dihydrogen phosphate,1.0 g sodi-um dodecyl sulfate,1 ml glacial acetic acid dissolved to 1 000 ml)(65∶35,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 275 nm,column temperature was 25 ℃,and the injection volume 20 μl. RESULTS:The linear range was 500-4 500 ng for hesperidin(r=0.999 8),38.816-388.16 ng for nobiletin(r=0.999 6),19.936-199.36 ng for tangeretin(r=0.999 5)and 640-2 560 ng for synephrine(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 96.42%-102.75%(RSD=2.54%,n=6),97.42%-99.95%(RSD=2.46%,n=6),99.26%-106.19%(RSD=2.31%,n=6) and 97.47%-99.76%(RSD=1.95%,n=6),respectively. CONCLUSIONS:The method is simple and stable with good reproducibility, and can be used for the contents determination of main active components in C. chacheiensis with different storage time. Pericarpi-um citri“the older the better”may be irrelevant to the change of the contents of the above-mentioned 4 active components,and it is speculated related to the release of volatile oil content to ease dryness.

The treatment of cerebral palsy is still a big medical problem.The pathogenesis of spasticity,as result of a variety of lesions of the cerebral cortex,brain stem,spinal cord,is caused by involvement of the inhibitory pyramidal and parapyramidal descending tracts terminating on the spinal facilitatory myotatic reflex.A lesion of the descending tracts disturbs this equilibrium leading to spasticity,which is cha-racterized by muscle resistance at rest that is velocity dependent and associated with an increase in tonic stretch reflexes resulting from hype-rexcitability of the stretch reflex.Spasticity caused abnomal posture,caused special movement,and higher multilation,which affect children's life severely.There are so many ways to lower hypermyotonia,such as:drugs,rehabilitation care,acupuncture and so on.Bioelectric stimulation therapy is a new ways in the zone.Its curative effect and the mechanism are still in explore,this article just to give an overview about bioelectric stimulation therapy in curing spastic cerebral palsy.

AIM: Currently available anti-scarring regimens for glaucoma filtration surgery have potentially blinding complications and safer alternatives would be beneficial. This experiment is to investigate the effect of TGF-β2 antisense oligodeoxynucleotide on differentiation, proliferation of subconjunctival fibroblast following glaucoma filtration surgery.METHODS: Glaucoma filtration surgery were performed on both eyes of 28 rabbits. TGF-β2 antisense oligodeoxynudeotide was subconjunctivally injected in the right eyes (A group), and TGF-β2 missense oligodeoxynucleotide (B group)or PBS(C group) was used at the same method in the left eyes as controls. Rabbits were killed at 4,7,14 and 28 days after surgery. Intraocular pressure (IOP), bleb characteristics were recorded at different time point. Subconjunctival fibroblasts were examined by immunohistochemistry and electron microscopy.RESULTS: The IOP of rabbits in group A was significantly lower at 14 days (6.74± 1.18 mmHg) and 21 days (8.15± 1.97mmHg) after operation than the IOP in group B (8.53± 1.04,9.72± 1.09 mmHg)(P ＜0.01) and group C(8.79± 1.21, 9.43±1.27 mmHg) (P ＜0.05). The mean bleb survival time was longer (17.2 days) in group A than that of group B (14.5 days) and group C (13.5 days)(P＜0.05). The population of the cells expressing α -smooth muscle actin(α -SMA) and proliferating cell nuclear antigen (PCNA) was significantly reduced in group A compared with the group B and C. The ultrastructure of fibroblast was not altered by TGF-β2 anti-sense oligodeoxynucleotide.CONCLUSION:TGF-β2 antisense oligodeoxynucleotide can prevent the scar formation after glaucoma surgery by inhibit the differentiation and proliferation of subconjunctival fibroblast. It could be a potentially useful anti-scarring alternative for the prevention of late surgical failure.

0.05).The level of TG,TC and LDL-C of high cholesterol group was higher than that of the other groups(P

OBJECTIVE:To establish the quality standard of Oleanolic acid dripping pills. METHODS:The property of the preparation was identified,and weight difference and dissolution time limit were detected. UPLC method was adopted to determine the content of oleanolic acid in the preparation. The determination was performed on ACQUITY UPLC BEH C18 column with mo-bile phase consisted of acetonitrile-water (70:30,V/V) at the flow rate of 0.30 mL/min. The detection wavelength was set at 206 nm,the column temperature was 30 ℃,and the sample was 5 μL. RESULTS:The characteristics of the preparation was significant;weight difference ranged 37.62%-46.56%;dissolution time limit was 24 min. Linear range of oleanolic acid ranged 0.006-0.06 mg/mL (r=0.9998). RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recoveries were 99.34%-100.40%(RSD=0.4%,n=6). CONCLUSIONS:Established standard can be used for quality control of Oleanolic acid dripping pills.