Objective To prepare and characterize nanoparticles in-loaded one-way release-controlled chitosan membrane,and to explore the release-controlled rule of the film in vitro.Methods The chitosan nanoparticles were prepared by inverse crosslinking-emulsion method.The one-way release-controlled membrane was prepared by a casting method.Transmission electron microscope (TEM) was used to evaluate the morphological properties and particle size analyzer was used to analyze particle size distribution.The morphology of the membrane was inspected through scanning electron microscope (SEM).MTT assay was applied to determine the biological safety of chitosan nanoparticles.The distribution of the nanoparticles was observed by fluorescence microscope.The in vitro release studies were adopted to evaluate the release-controlled rule.Results The four kinds of nanoparticles had spherical shapes and uniform particle size.The size of the hyaluronic acid-coated chitosan nanoparticle was (255.40±39.10) nm.Hyaluronic acid-coated chitosan nanoparticles showed the best property of sustained release and biocompatibility.The membrane had a loose inner layer and a dense outer layer,and the distribution of the nanoparticles was uniform in the inner layer of the membrane.The release of protein from membrane was unidirectional and the membrane displayed good controlled release property.Conclusions The nanoparticles in-loaded one-way release-controlled chitosan membrane presents good one-way sustained release performance.It is potentially useful in delivery system of growth factors.

Objective To investigate the effect,security and tolerance of pregabalin combined with sertraline hydrochloride for refractory neuropathic pain.Methods Forty-six patients with refractory neuropathic pain were divided into two groups by random digits table.The patients in pregabalin group (24 patients) were treated by pregabalin combined with sertraline hydrochloride,and the patients in carbamazepine group (22 patients) were treated by carbamazepine combined with sertraline hydrochloride.They were treated for 4 weeks.The scores of visual analogue scales (VAS),Hamilton rating scale of depression (HAMD),Hamilton rating scale for anxiety (HAMA) and the dose,adverse reaction were compared between two groups.Results The total effective rate of pregabalin group was 83.3％(20/24) and carbamazepine group was 86.4％ (19/22),there was no significant difference (P＞ 0.05).After treatment of 1,2,4 weeks,the scores of VAS in two groups were significantly decreased compared with those before treatment[pregabalin group:(5.90 ± 1.11),(4.59 ± 1.33),(3.59 ± 1.94) scores vs.(6.50 ± 0.85) scores; carbamazepine group:(5.50 ± 1.06),(4.18 ± 1.71),(3.73 ± 1.88) scores vs.(6.50 ± 0.80) scores] (P ＜0.05).After treatment of 2 weeks,the scores of VAS in carbamazepine group was significantly lower than that in pregabalin group (P ＜ 0.05).In pregabalin group,the scores of HAMD/HAMA after treatment of 1 week had no significant difference compared with before treatment (P ＞ 0.05),and the scores of HAMD/HAMA after treatment of 2,4 weeks were significantly increased [(15.25 ± 4.15),(11.29 ± 4.16)scores vs.(16.08 ± 3.56) scores and (14.21 ± 4.72),(11.63 ± 4.50) scores vs.(15.25 ± 4.05) scores] (P ＜0.05).In carbamazepine group,the scores of HAMD/HAMA after treatment of 1,2 weeks was decreased compared with those before treatment,but there was no significant difference (P ＞ 0.05).The scores of HAMD/HAMA after treatment of 4 weeks was significantly decreased [(13.18 ± 2.42) scores vs.(15.20 ±1.05) scores,(12.32 ± 2.64) scores vs.(14.55 ± 4.43) scores] (P ＜ 0.05).The scores of HAMD/HAMA in pregabalin group after treatment of 4 weeks was significantly lower than that in carbamazepine group (P ＜0.05).There was no severe adverse reaction.Conclusion Pregabalin combined with sertraline hydrochloride is worth to recommend for refractory neuropathic pain,and has no serious adverse reaction.

At present, antibacterial drugs are widely used in the clinical treatment of infectious diseases. It is particularly impor-tant to focus on the safety of antibacterial drugs for the application improvement in the clinical treatment. The paper reviewed and sys-tematically analyzed the relative literatures in order to explain the pathomechanism of drug-induced renal injury caused by antibacterial drugs and propose some preventive measures. The study suggested that attention should be paid to the distribution and characteristics of the adverse drug reaction of antibacterial drugs to ensure the safe and proper administration of the drugs.

Objective To prepare three kinds of biodegradable film materials used for protein drug carrier,and compare their degradation and drug release behavior.Methods Three different biodegradable and controlled release films,gelatin,chitosan oligosaccharides and crosslinked chitosan oligosaccharides films were prepared.Protein release behavior was determined by the Bradford.At the same time,degradation rate and swelling rate were tested,and the biocompatibility of film was investigated by MTT assay.Results The release time of crosslinked chitosan oligosaccharides film was 168 h,which was longer than that of chitosan oligosaccharides film,and different in different solution.The degradation rate and the swelling rate of crosslinked chitosan oligosaccharides was 60％ (360 h) and 110.45％,respectively,while the chitosan oligosaccharides membrane was 80％ (360 h) and 113.03％.The MTI assay revealed that the crosslinked chitosan oligosaccharides film had better biocompatibility.Conclusions By comparing different properties,the crosslinked chitosan oligosaccharides film is the best choice for protein drug carrier.

Background Our previous study demonstrated that epigallocateehin-gallate(EGCG),an active ingredient of green tea,has protective effect on optical nerve after optic nerve crush.Astrocyte was proved to play key role in the repair of nerve tissue,but the influence of EGCG on astrocyte is unclear.Glial flbrillary acidic protein (GFAP) is a special marker for astrocyte. Objective The aim of this study was to investigate the effect of EGCG on the expression of GFAP in optic nerve tissue after optic nerve crush. Methods Seventy-two clean Wistar rats were randomly divided into normal control group,sham+EGCG group,optic nerve crush+normal saline group(vehicle group),optic nerve crush+EGCG group.Optic nerve crush models were established by clamping optical nerve for 60 seconds by minitype optic nerve clipper with the force of 40 gram.Only ocular tissue was cut in the rats in sham group.Normal saline solution or EGCG(25 mg/kg)was intraperitoneally injected daily for 5 days consecutively and orally administered(2 mg/kg)daily afterwards.The expression of GFAP in optic nerve was detected by immunohistochemistry and quantified by Western blotting analysis on day 7,14 and 28 after modeling. Results lmmunochemistry showed that GFAP were weakly expressed in the rats of both normal group and sham+EGCG group with the sliSht brown staining in optic nerve tissue.The deeply brown staining for GFAP was seen in vehicle group,and the staining intensity weakened in optic nerve crush+EGCG group compared to vehicle group on days 7,14 and 28 after modeling.Western blotting analysis revealed that the expression level of GFAP in rat optic nerve tissue of vehicle group was significantly enhanced in comparison with normal control group(P＜0.01).On day 7 and 14 after optic nerve modeling,the expression levels of GFAP were evidently decreased in optic nerve crush+EGCG group in comparison with vehicle group(P＜0.05).However,on day 28 after modeling,no significant difference wag found in the expression levels of GFAP between vehicle group and optic nerve erush+EGCG group(P＞0.05). Conclusion EGCG down-regulates optic nerve crush-induced of GFAP in the optic nerve and therefore attenuates the activity of astrocytes,suggesting that EGCG might reduce the formation of glial scar.

Background Researches demonstrated that epigallocatechin-3-gallate(EGCG) can protect retinal ganglion cells(RGCs) against damage induced by retinal ischemia-reperfusion and optic nerve crush(ONC),but the effect of EGCG on lateral geniculate nucleus(LGN) was under study.Objective This study was designed to detect neuroprotective effect of EGCG on LGN in the rat model with ONC.Methods Forty-eight 7-week-old female clean Wistar rats were randomly divided into normal control group,sham operation+EGCG group,ONC+normal saline(NS) group and ONC+EGCG group.ONC models were created by clamping the optical nerve for 60 seconds with the clipper with the force of 40 grams in the right eyes of 24 rats.The EGCG solution(25mg/kg) was intraperitoneally injected from 2 days before operation daily for 5 consecutively days and orally administered(2mg/kg) after that,and NS was used in the same way for ONC+NS group.Four weeks after ONC,the brain tissue of the rats was obtained,and the neurons of dorsal LGN(dLGN) were counted by Nissl staining under the light microscopy.The expression of neurofilament triplet L(NF-L) was detected by immunohistochemistry and Western blot analysis.Meanwhile,the neuronal nitric oxide synthase(nNOS) positive cells were counted.Results Compared with normal control group,no significant differences were found in neuron number both between sham operation+EGCG group or ipsilateral LGN of operative eyes in ONC+normal saline group and ONC+EGCG group(P=0.906,0.561,0.794,0.646 respectively) in 4 weeks after ONC,but loss of neurons in contralateral LGN in both ONC+normal saline group and ONC+EGCG group were observed(P=0.000,0.015 respectively).However,compared with ONC+normal saline group,the density of neurons in ONC+EGCG group was higher(P=0.007).Moreover,a higher expression level of NF-L protein was seen in ONC+EGCG group compared with ONC+normal saline group at contralateral LGN of operative eyes(P=0.002).Concerning the number of nNOS positive cells in LGN,there was no significant difference among normal control group,sham operation+EGCG group and ONC+EGCG group(P＞0.05).The number of nNOS positive cells in the contralateral LGN of operative eyes of ONC+normal saline group was higher than that of ONC+EGCG group(P=0.000).Conclusion EGCG plays the protective effect on LGN after ONC in rats through mediating the expression of nNOS.

Background The underlying mechanism of choroidal neovascularization(CNV) is multifactorial and complex.Focal adhesion kinase(FAK) plays a crucial role in controlling essential cellular processes and influencing distinct steps of the angiogenic response.But to our knowledge,seldom study on the effect of FAK on CNV formation has been reported previously.Objective In this study,the effect of several CNV risk factors on the expression of FAK in cultured retinal pigment epithelium(RPE) cells was investigated to illuminate effect of FAK on CNV.Methods Human RPE cells were isolated from donor eyes and exposed to H2O2,swallow of outer segment of photoreceptors(POS) and extracellular matrix(ECM) separately with the treating as follows:RPE cells were co-cultured with 10,20,50 and 100μmol/L H2O2 for 20 days;POS(1×106/ml) were co-cultivated with RPE cells for 20 days(setting control group,POS group,hypoxia group with 200μmol/L CoCl2,and POS+hyoxia group);RPE cells were cultured on the plates coated with 100mg/L fibronectin(FN),laminin(LN) or collagen typeⅠfor 30minutes or 1 hour.The expression of FAK and pFAK in RPE cells were examined by Western blot analysis.Results FAK was highly expressed in the 20μmol/L and 50μmol/L H2O2 groups compared with control group(P＜0.01);while he expression level of pFAK was reduced after treated with H2O2 in comparison with the control group(P＜0.01).After cultured with POS for 20 days,the undigested lysosome could be observed in RPE cells.The expressions of FAK and pFAK in RPE cells were not significantly changed between control group and POS groups(P＞0.05),but those in hypoxia group were significantly up-regulated in comparison with control group(P＜0.01).Compared with the hypoxia group,the expression amount of pFAK was elevated in POS+hyoxia group(P＜0.01).In comparison with control group,the increased pFAK expression was seen in FN,LN and collagen typeⅠtreating for 1-hour groups(P＜0.05,P＜0.01).Conclusion FAK pathway participates in several CNV-initiated signaling,such as H2O2,POS and ECM,in cultured RPE cells.It is reasonable to believe that FAK potentially plays an important role in CNV-dependent disorder.

Objective To search for the changes of T cells subpopulations and immunoglobulin and their relation-ship in children patients with simple nephrotic syndrome. Design Case-control research. Patients aud Participants 39 patients with simple nephrotic syndrome were divided into two groups:the incipient group and relapse group (6 cases were determined at the incipient and relapse time) .Thereare 28 patients in incipient group, 19 males and 9 females, at the age of 2 to 10 years old. There are20 patients in relapse group, 12 males and 8 females, at the age of 3 to 13 years old. There are 35health children in control group, 21 males and 14 females, 2～13 years old. Interventions T cells subpopulations were determined by indirect immunofluorescence of OKT linesmonoclonal antibodies. The serum IgG was determined by routine simple agar immunodiffusion tests. Results and Conclusions The CD_3~+ and CD_4~+ cells are of no change in the children patients withsimple nephrotic syndrome, and the CD_8~+ and CD_(10)~+ cells are obviously increased, the Values of CD_4~+/CD_8~+ are obviously lower than those in the control qroup, there are no difference between the incipientand relapse groups. The levels of serum IgG were decreased in the 85.3% children patients, IgM were inc-reased in 29.4% of that. The values of CD_4~+/CD_8~+ have positive correlation and negative correlationwith the levels of serum IgG and IgM respectively.

OBJECTIVETo investigate the potential protective effect of the mitochondria-targeted antioxidant Mitoquinone (MitoQ) on post-thaw human sperm.

METHODSSemen samples were collected from 60 normal fertile men, each divided into six parts of equal volume to be incubated at 37 °C in normal saline (G0, control) or in the extender with 2 nmol/L (G1), 20 nmol/L (G2), 200 nmol/L (G3), 2 µmol/L (G4), and 20 µmol/L of MitoQ (G5). After one hour of incubation, the samples were subjected to computer-assisted semen analysis (CASA) for sperm motility, flow cytometry for reactive oxygen species (ROS), thiobarbituric acid assay for the concentration of malondialdehyde (MDA), and MitoTracker fluorescent staining and flow cytometry for the sperm mitochondrial membrane potential (MMP). Then, the semen were cryopreserved with none (B0), 200 nmol/L (B1), and 2 µmol/L of MitoQ (B2), followed by detection of the changes in the ROS, MDA, and MMP of the post-thaw sperm.

RESULTSThe percentage of progressively motile sperm and total rate of sperm motility were significantly higher in G3 ([30.8 ± 10.2]% and [70.6 ± 9.0]%) and G4 ([32.7 ± 13.5]% and [70.3 ± 11.9]%) than in G0 ([17.6 ± 5.0]% and [54.9 ± 11.5]%) (P < 0.05). The level of ROS dropped markedly with the increased concentration of MitoQ, 86.5 ± 31.6 in G3, 93.6 ± 42.0 in G4, and 45.1 ± 15.0 in G5, as compared with 160.8 ± 39.7 in G0 (P < 0.05). The content of MDA was remarkably lower in G3 ([0.9 ± 0.5] µmol/mg) and G4 ([0.9 ± 0.5] µmol/mg) than in G0 ([1.9 ± 1.1] µmol/mg) (P < 0.05), but not in G5 ([1.7 ± 0.7] µmol/mg), which was even higher than in G3 and G4 (P < 0.05). The MMP showed a significant reduction in G5 (1156 ± 216) in comparison with G0 (1701 ± 251) (P < 0.05) but exhibited no remarkable difference between G0 and G1 (1810 ± 298), G2 (1995 ± 437), G3 (1950 ± 334), or G4 (1582 ± 314). The percentage of progressively motile sperm and total rate of sperm motility after freezing-thawing were significantly decreased as compared with those of the fresh semen (P < 0.01), but both were remarkably higher in B1 ([3.2 ± 2.3]% and [ 43.0 ± 9.5]%) than in B0 ([0.8 ± 0.6]% and [26.5 ± 11.4]%) (P < 0.05). The ROS level was significantly lower in B1 and B2 than in B0 (34.6 ± 12. 3 and 37.0 ± 10.5 vs 56.9 ± 14.3, P < 0.05), and so was the MDA content ([1.4 ± 0.5] and [1.4 ± 0.6] µmol/mg vs [2.6 ± 1.0] µmol/mg, P < 0.05), but the MMP was markedly higher in B1 and B2 than in B0 (1010.0 ± 130.5 and 880.6 ± 128.6 vs 721.1 ± 24.8, P < 0.05).

CONCLUSIONAddition of MitoQ to the freezing extender at 200 nmol/L may effectively improve the quality of human sperm and MitoQ is a good protective addictive for human sperm cryopreservation.

Antioxidants ; Cryopreservation ; Humans ; Male ; Malondialdehyde ; analysis ; Membrane Potential, Mitochondrial ; Mitochondria ; Organophosphorus Compounds ; pharmacology ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; Semen ; Semen Analysis ; Semen Preservation ; Sperm Motility ; Spermatozoa ; drug effects ; Ubiquinone ; analogs & derivatives ; pharmacology