Background Nearly over 40 years have elapsed since the original findings of visual cortical plasticity,but none of drug has been found for curing amblyopia effectively. Objective The goal of this study was to investigate the effects of different dose of levodopa on flash visual evoked potential(F-VEP)and morphology of visual cortex cells in monocular deprivation rat and explore the possible mechanism of curing amblyopia.Methods Monocular deprivation model were established by suturing eyelids of 30 2-week-old Sprague Dawley(SD)rats for 4 weeks.The 30 SD rats were then divided into 3 groups randomly and 10 rats for each group.Normal saline.20 ms/kg levodopa,80 ms/kg levodopa were intragastrically administered once per day after modeling respectively for 4 weeks.F-VEP was recorded after establishment of model and administration of drug respectively.The rats were sacrificed and the visual cogex was obtained for histological examination,and TUNEL technique was used to assess the structural change of visual cortex.Results The latency of P1 wave was significantly longer in the deprived eye than the normal eyes(P<0.05).After administration of levodopa,the latent periods of Pl wave in the deprived eye were obviously shortened in comparison with before administration of levodopa in 20 ms/kg and 80 mg/kg levodopa group (P<0.05).The difference values of latent period of P1 wave between before and after administration of drug showed statistically significant change in three groups(P<0.05).No evidently alterations were found in the amplitude differences of N1 P1 and P1 N2 waves among three groups(P>0.05).The number and structure of neurons in contralateral visual cortex of non-deprived eye were normal.However,the numbers of neurons in deprived eye were significantly less and presented the signs of para-apoptosis in normal saline group.In 20 mg/kg levodopa groups,the alterations of number and morphology in neurons of rat visual eogex were slight.TUNEL assay revealed that the numbers of positive neurons in contralateral visual codex of non-deprived eye were 2.20±1.23.while those in deprived eye were 53.7±9.36,27.20 4±5.96 and 10.70±3.23 in normal saline group,20 mg/kg and 80 mg/kg levodopa group respectively,showing a significant difference among them(P>0.05).After usage of levodopa,the numbers of positive neurons was negatively correlated with the difference value of P,latent period of VEP(r=-0.815,P=0.000).Conclusion Levodopa has a therapeutic effect on rat deprived eye,and its possible mechanism is inhibiting the para-apoptosis of neurons and participating in the development and plasticity of visual system.

OBJECTIVE: To establish a multiplex STR genotyping method for autosomal STR and Y-STR loci in forensic biological practice. METHODS: Widely used autosomal STR loci and Y-STR loci were selected. A set of PCR primers was designed, and a 5-dye fluorescent labeled STR multiplex PCR reagent kit was developed. RESULTS: A kit was developed which can simultaneously detect 15 autosomal STR loci, 10 Y-STR loci, and an Amelogenin. CONCLUSION The 15 autosomal STR plus 10 Y-STR kit in combination with capillary electrophoresis method was used to STR genotyping with accurate and reliable results. The new one-step testing kit can potentially be widely used in forensic cases and DNA databank in the future.
Amelogenin ; Chromosomes, Human, Y/genetics* ; DNA Primers ; Databases, Nucleic Acid ; Forensic Genetics/methods* ; Genotype ; Genotyping Techniques/instrumentation* ; Humans ; Indicators and Reagents ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction

Phytantriol (PT), ethanol (ET) and water were used to prepare in situ cubic liquid crystal (ISV2). The pseudo-ternary phase diagram of PT-ET-water was constructed and isotropic solution formulations were chosen for further optimization. The physicochemical properties of isotropic solution formulations were evaluated to optimize the composition of ISV2. In situ hexagonal liquid crystals (ISH2) were prepared based on the composition of ISV2 with the addition of vitamin E acetate (VitEA) and the amount of VitEA was optimized by in vitro release behavior. The phase structures of liquid crystalline gels formed by ISV2 and ISH2 in excess water were confirmed by crossed polarized light microscopy and small angle X-ray scattering, respectively. Rheological properties of ISV2 and ISH2 were studied by a DHR-2 rheometer. In vitro drug release studies were conducted by using a dialysis membrane diffusion method. Pharmacokinetics was investigated by determination of sinomenine hydrochloride (SMH) concentration in synovial membrane after intra-articular injection of SMH-loaded ISH2 in adjuvant-induced arthritis rats. The optimal ISV2 (PT/ET/water, 64 : 16 : 20, w/w/w) loaded with 6 mg x g(-1) of SMH showed a suitable pH, injectable and formed a cubic liquid crystalline gel in situ with minimum water absorption in the shortest time. The optimal ISV2 was able to sustain the drug release for 144 h. The optimal ISH2 system was prepared by addition of 5% VitEA into PT in the optimal ISV2 system. This ISH2 (PT/VitEA/ET/water, 60.8 : 3.2 : 16 : 20, w/w/w/w) was an injectable isotropic solution with suitable pH. The new ISH2 was able to sustain the drug release for more than 240 h. Local pharmacokinetics study indicated that the retention time and AUC(0-∞) of ISH2 group were increased significantly compared with that of SMH solution group and the AUC(0-∞) of ISH2 group was 6.01 times higher than that of SMH solution group. The developed ISH2 was suitable for intra-articular injection that may apply to patients in the treatment of rheumatoid arthritis.
Animals ; Chemistry, Pharmaceutical ; Diffusion ; Ethanol ; Fatty Alcohols ; Gels ; Injections, Intra-Articular ; Liquid Crystals ; Morphinans ; administration & dosage ; chemistry ; Rats ; Rheology ; Water ; alpha-Tocopherol

OBJECTIVETo observe the effect of Astragalus Injection (ASI) combined with rat's mesenchymal stem cells transplantation (rMSCs) for repairing spinal cord injury in rats.

METHODSOne hundred and twenty Wistar rats were randomly divided into 6 groups, named respectively by letters from A-F. they were treated respectively with none, PBS solution, ASI, rMSCs, and ASI + rMSCs, with the ASI administered via intraperitoneal injection and the rMSCs given by local injection to the spinal cord, on the 3rd day of operation. The condition of nerve function recovery was assessed on the 7th, 14th, 21st and 28th day after treatment by scoring according to the Basso-Beattie-Bresnahan (BBB) locomotor rating scale. Besides, the pathological change of the injured spinal cord was observed and expressions of glial fibrillary acidic protein (GFAP) and neurofilament-M (NF-M) in the BrdU-labeled rMSCs in the spinal cord tissue were examined by immune histochemistry.

RESULTSThe recovery of spinal cord in Group D, E and F was better than that in Group B and C, showing higher BBB scores. As compared with Group E, Group F showed a higher score of nerve function and a milder inflammatory cell infiltration with lessened tissue edema in the spinal cord and more active proliferation of gliacyte. Double-labelled immunohistochemical examination showed that the transplanted rMSCs were alive in the host's spinal cord, revealing the expressions of GFAP and NF-M from the 7th day after transplantation, which were migrating to the injured site. The amount of GFAP and NF-M positive cells in Group F was much more than that in Group E (P < 0.05)

CONCLUSIONTransplantation of rMSCs is an effective method in treatment of the spinal cord injury; ASI has the capacity for inducing rMSCs differentiated into neurons, and could synergize with rMSCs to promote the repairing of the spinal cord from injury.

Animals ; Astragalus Plant ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Mesenchymal Stem Cell Transplantation ; Nerve Regeneration ; Phytotherapy ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; therapy

Objective:To investigate the relationship between the expression of serum exsomal miRNA-155-5p (miR-155-5p) and prognosis in patients with esophageal squamous cell carcinoma (ESCC).Methods:A total of 336 samples from ESCC patients in Fujian Provincial Cancer Hospital from October 2014 to December 2015 were collected. The relative expression levels of serum exsomal miR-155-5p were detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). Cut-off value of the expression levels of serum exsomal miR-155-5p was determined by using X-tile software. Based on the optimal cut-off value, patients were divided into miR-155-5p low expression group and miR-155-5p high expression group. The survival curve was drawn by using Kaplan-Meier method and Cox proportional hazards regression model was used to make survival analysis.Results:The cut-off value of serum exsomal miR-155-5p expression level was 2.340. According to the cut-off value, patients were divided into miR-155-5p low expression group (<2.340) of 51 cases and miR-155-5p high expression group (≥2.340) of 285 cases. There were no statistical differences in age ( χ2 = 0.020, P = 0.887), gender ( χ2 = 0.283, P = 0.595), tumor location ( χ2 = 0.063, P = 0.977), differentiation grade ( P = 0.474), clinical staging ( χ2 = 3.996, P = 0.136) and surgery treatment ( χ2 = 0.941, P = 0.332) of patients in both groups. ESCC patients in serum exsomal miR-155-5p high expression had a higher risk of death compared with patients in miR-155-5p low expression group ( HR = 1.763, 95% CI 1.049-2.961, P = 0.032). Conclusion:The high expression level of serum exsomal miR-155-5p is associated with poor prognosis in ESCC patients and it could be used as a prognostic new marker in ESCC patients.

AIM and METHODS: To study the culture met ho d of human neural stem/progenitor cells, these cells derived from human fetal fo rebrains were maintained and expanded in serum-free defined medium containing ba sic fibroblast growth factors (bFGF), epidermal growth factor (EGF) and B27. Whe n they formed neurosphere, these three factors and supplemented FBS were removed to induce differentiation. Cell were cultured for 12-14 d, then fixed for immun ocytochemistry examination. RESULTS: This period of expansion resulted in a 107-fold incre ase in this heterogeneous population of cells. Upon differentiation, they form n eurons, astrocytes and oligodendrocytes, the three main phenotypes in the CNS. CONCLUSION: These results demonstrate the feasibility of long-t erm in vitro expansion of human neural progenitor cells. The advantages of s uch a population of neural precursors for allogeneic transplantation, including t he ability to provide an expandable, well-characterized, defined cell source, ca n form specific neuronal or glial subtypes.

This study was conducted to investigate the paclitaxel loaded by hydrazone bonds in poly(ethylene glycol)-poly(caprolactone) micelles (mPEG-PCL-PTX) on proliferation and apoptosis of human lung cancer A549 cells and its possible mechanisms of anti-tumor activity. The cell proliferation was measured with MTT assay. Flow cytometry were used to analyze the cell cycle. The cell apoptosis was analyzed using Hoechst/P staining. The expression levels of apoptotic genes expression in the mitochondrial apoptosis pathway were detected by RT-PCR and Western blotting, respectively. The mPEG-PCL-PTX could inhibit the proliferation of A549 cells and promote the apoptosis. The Bax, caspase-3 protein expression were increased while Bcl-2 protein expression was decreased in A549 cells. Results showed that the polymer containing hydrazone bond is non-toxic in vitro, the mPEG-PCL-PTX micelles can inhibit the proliferation and induce the apoptosis of A549 cells. Key words: paclitaxel; micelle; A549 cell; proliferation; cell cycle; apoptosis
Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Micelles ; Paclitaxel ; pharmacology ; Polyesters ; Polyethylene Glycols ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

ObjectiveTo investigate the transcriptional changes of nitochondria fission and fusion gene loci and reactive oxygen species (ROS) level in cortical neurons of rats with chronic fluorosis,and to reveal their roles in mitochondria damage due to chronic fluorosis.MethodsSD rats were fed with different doses of fluoride through drinking water[＜ 0.5(control),10,50 mg/L,respectively] for 3 and 6 months.The level of ROS and mRNA contents of mitochondria fission gene loci Drp1/Fis1 and fusion gene locus Mfn1 in the cortical neurons of rat brains were detected with ROS fluorescent probe and real-time PCR,respectively.ResultsAs compared with control group [10.43 ± 5.98,(3.4 ± 0.6) × 103,(8.8 ± 1.4) × 10,(1.2 ± 0.2) × 102] at the experiment period of 3 months,the level of ROS and mRNA contents of mitochondria fusion gene locus Mfn1 and fission gene loci Drp1/Fis1 in the cortical neurons were obviously increased in the rats fed with 50 mg/L fluoride through drinking water[25.48 ± 6.09,(1.0 ± 0.2) × 1011,(3.0 ± 1.6) × 103,(8.9 ± 3.6) × 102,all P ＜ 0.05],whereas no significant changes were found in the rats fed with 10 mg/L fluoride[11.67 ± 3.49,(3.1 ± 0.3) × 104,(6.7 ± 2.7) × 10,(5.0 ± 0.9) × 10,all P ＞0.05].Furthermore,at 6 months of the experiment the increases in ROS level(63.02 ± 8.15,65.60 ± 7.40) and mRNA contents of mitochondria fission gene loci Drp1/Fis1 [(2.0 ± 0.8) × 106,(4.0 ± 0.6) × 105,(3.8 ± 1.3) × 103,(1.3 ± 0.2) × 103] and the decrease in mitochondrial fusion gene locus Mfn1[(3.0 ± 0.4) × 106、(4.0 ± 0.9) × 104]were observed in the cortical neurons of the rats fed with 10 mg/L and 50 mg/L fluoride as compared with the control group[25.26 ± 6.41,(3.0 ± 0.8) × 109,(5.1 ± 0.8) × 103,(2.8 ± 0.7) × 102,all P ＜ 0.05].Conclusions Excessive intake of fluorine leads to elevated ROS levels,and decreased transcription of mitochondrial fusion gene loci Mfn1,which is positively correlated with the time and dose-exposed to fluoride.The changes of mitochondrial fission and fusion gene loci in the cortical neurons may be related to high level of oxidative stress induced by chronic fluorosis.

Effects of dihydroartemisinin (DHA) on the expression level of Alpha-7 .3 giardin mRNA in C2 Giardia lam-blia was investigated in this study to explore the damage to skeleton protein of C 2 Giardia lamblia .Giardia lamblia was culti-vated respectively for 2 ,4 ,8 ,and 12 hours with modified TYI-S-33 medium containing 100 μg/mL and 200 μg/mL DHA , while the control group performed in the same experimental conditions without DHA .The expressive quantity of Alpha-7 .3 gi-ardin mRNA was determined by using real-time reverse transcription PCR ,and then we found that the expressive quantities of Alpha-7 .3 giardin mRNA with DHA were significantly lower than those in the control group .It’s suggested that dihydroarte-misinin has obvious inhibitory effect on the expression level of Alpha-7 .3 giardin mRNA in C2 Giardia lamblia .The actions of dihydroartemisinin on skeleton protein of C2 Giardia lamblia are effective .

Objective To investigate anorectal infection with Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) among men who have sex with men (MSM) with anorectal condyloma acuminatum(CA),and provide evidence for the clinical diagnosis and treatment of anorectal CA.Methods Anorectal swabs were obtained from MSM who were diagnosed with anorectal CA(trial group) and without CA(control group),nucleic acid fragments of NG and CT were detected by fluorescent quantitative polymerase chain reaction.Data about social demographic characteristics and sexual behaviors of investigated people were collected,compared and analyzed.Results A total of 158 patients were enrolled in this study,63 were in trial group and 95 in control group.Among 63 patients in trial group,infection rate of NG and CT were 17.46％ and 28.57％ respectively,co infection rate of NG and CT was 12.70％;in control group,infection rate of NG and CT were 6.32 ％ and 9.47 ％ respectively,co-infection rate of NG and CT was 2.16％;infection rate of NG and CT,as well as co-infection rate of NG and CT in trial group were all significantly higher than control group(all P＜0.05).Conclusion Infection rate of NG and CT is high in MSM with anorectal CA,suggesting that more attention should be paid to the screening of NG,CT and other sexually transmitted infection among those who were clinically diagnosed with anorectal CA.