Background Marrow mesenchymal stem cells (MSCs) has been successful induced to differentiate into corneal cells,retinal ganglion cells (RGCs) and retinal neuron-like cells in recent years,but there are few studies about MSCs induced into photoreceptor cells and their microenvironment.Objective The aim of this study was to explore the induce and differentiation of bone marrow mesenchymal stem cells (BMSCs) into photoreceptor-like cells in vitro and microenvironment.Methods The second generation of human BMSCs strain and RPE cells strain were cultured and passaged,respectively,and the fourth generation of BMSCs and the third generation of RPE cells were used in the experiment.BMSCs were cocultured using the mesenchymal stem cells medium (MSCM) contained 20 μg/L basic fibroblast growth factor (bFGF),20 μg/L epithelial growth factor (EGF)and 20 μg/L brain derived neurotrophic factor (BDNF) with RPE cells to induce the differentiation of BMSCs in the induced group,and BMSCs were cultured in MSCM only in the control group.The morphology of induced and differentiated cells were observed under the inverted microscope.Inmmunocytochemistry was used in induced for 3-,5-,7-day cells to detect the expression rate of rhodopsin protein for the identification of phenotype of the differentiated cells.RT-PCR was used in induced for 5-,7-day cells to detect the expressions of rhodopsin mRNA and recoverin mRNA.Results Cultured BMSCs grew well with the spindle shape,and passaged RPE cells showed the uniform size and polygon shape with the abundant pigment in the cells.Some induced cells appeared to be the neuron-like cells with round shape and long prominence and the secondary reticular branches.The expression rates of rhodopsinin the cells were (5.83±0.29)％,(20.36±0.32)％ and (29.80±2.30)％ in the third,fifth and seventh day after induce,which were significantly higher than (0.71 ±0.35) ％,(2.56±0.24) ％ and (2.32±0.42) ％ of control cells (t3 d =41.510,t5d =107.290,t7 d =30.036,P＜0.01).The grey scales of rhodopsin mRNA and recoverin mRNA were significantly elevated in the induced and differentiated cells compared with control cells in the fifth and seventh day (rhodopsin mRNA:t5 d =103.506,t7 d =122.584,P＜0.01 ; recoverin mRNA:t5 d =106.674,t7 d =189.992,P＜0.01).Conclusions BMSCs can be successfully induced to differentiate into photoreceptor cells after cocultured by conditioned medium with RPE cells.

Anti-Infective Agents ; therapeutic use ; Bacterial Infections ; drug therapy ; Child ; Humans

AIM and METHODS: To observe the effects of glucose-free and Mg 2+ -free in the extracellular fluid on the changes of [Ca 2+ ] i in the cerebro-cortical neurons damaged by 1 mmol/L glutamate using laser confocal scanning microscope. RESULTS: Both frequency and amplitude of neuronal calcium oscillation induced by glutamate were lowered in glucose-free and Mg 2+ -free buffers. The basic [Ca 2+ ] i concentration was lowered in the former case , but it was elevated in the latter case. CONCLUSION: Mg 2+ -free aggravates [Ca 2+ ] i overload induced by 1 mmol/L glutamate ,under certain conditions the glucose-free might resist damage role of glutamate and Mg 2+ -free.

AIM: The purpose of the present study was to detect intracellular Ca 2+ changes in living brain slices during focal cerebral ischemia/reperfusion (I/R) and reveal the role of intracellular Ca 2+ in the cerebral I/R injury. METHODS: The model of focal cerebral I/R was established in rats by reversible inserting a nylon thread, and dynamic change of intracellular Ca 2+ in brain slices was determined using laser confocal imaging system. RESULTS: ① Ca 2+ gradually enhanced with increase in ischemic time in cortex and striatum. ②At 1 h ischemia/ 10 min reperfusion, Ca 2+ increased significantly in striatum, but Ca 2+ decreased at 3 h reperfusion compared with 10 min reperfusion. ③ Ca 2+ markedly enhanced at 6 h ischemia compared with 1 h ischemia, and after 3 h reperfusion Ca 2+ decreased, but was still higher than that in sham-operation group. ④The striatum is more sensitive than cortex to ischemia/reperfusion. CONCLUSION: Ca 2+ overload in the area of cortex and striatum may play an important role in cerebral ischemia/reperfusion injury in rats.

Objective To investigate the expression of transforming growth factor?1(TGF-?1)in human colorectal carcinoma and its value for predicting the prognosis.Methods The expression of TGF-?1 and vascular endothelial growth factor(VEGF)was measured in specimens of 52 coloreetal cancers by immunohistoehemistry.The features of clinical pathology were analyzed and the follow-up of all patients were conducted.The correlation between the expression of TGF-?1 and the survival time was studied with Log-rank test.Results Of 52 patients,no expression of TGF-?1 and VEGF was observed in 11 and 14 patients,and the expression was noticed in 41 and 38 patients,respectively.There was a signifi- cant positive correlation between expression of TGF-?1 and expression of VEGF(x~2＝0.633,P＜0.01). Furthermore,the expression of TGF-?1 was significantly correlated with Dukes staging(x~2＝19.866,P＜0.01)and metastasis of lymph nodes(x~2＝13.152,P＜0.01).The 3-year overall survival rates(OSR)in all patients was 49.1% and the 3-year OSR of patients with and without expression of TGF-?1 were 20.5% and 69.2% respectively(x~2＝11.64,P＝0.0006).Conclusion The expression of TGF-?1 could be served as an important predicator for prognosis of coloreetal carcinoma.

BackgroundRecent researches suggested that properties of neurons in the lateral geniculate neuron (LGN) may represent an important neural limitation on the development of basic spatial and temporal vision,and even binocular rivalry.However,previous studies on the properties of spatiotemporal frequency tuning of LGN were rather concentrated on a monkey or cat,whereas little is known about rat.ObjectiveThis study was to examine the development of spatiotemporal frequency tuning in rats LGN.MethodsTwenty Wistar rats were collected and divided into 14-16 day,20-22 day,27-30 day and 60 day groups according to the different ages after their eyes opened and 5 rats were assigned for each group.Extracellular single neuron recording was carried out in the rats to study the spatio-temporal receptive field properties of neurons in LGN by sinusoidal gratings visual stimuli.Dynamic changes of the spatio-temporal receptive field properties of neurons in LGN with the development of Wistar rats were evaluated.ResultsThe differences between band-pass and low-pass distribution of temporal frequency or spatial frequency of rat LGN were not statistically significant (x2 =0.68,0.47,P＞0.05 ).The optimal temporal frequency of receptive field in rat LGN went up to the maximum value until 60 day in Wistar rats.The mean optimal temporal frequencies of neurons in the four different age groups were ( 2.5 ± 1.3 ),( 2.6± 1.2),(2.6± 1.1 ) and ( 3.6± 1.1 ) Hz with significant differences among the 4 groups (F=4.53,P＜0.05 ),and those in the 14-16 day group,20-22 day group,27- 30 day group were significantly lower than in the 60 days group ( q =4.43,4.10,4.03,P ＜ 0.05 ).No significant differences were seen among the 14-16 day group,20-22 day group and 27-30 day group ( P＞0.05 ).The optimal spatial frequency values in the four groups were ( 0.04 ± 0.04 ),( 0.04 ± 0.03 ),( 0.05 ± 0.03 ),( 0.05 ± 0.04 ) cpd,respectively without statistical difference among them ( F =0.58,P ＞ 0.05 ).The temporal and spatial bandwidth values in the various age groups were not statistically significant among the four groups ( F =0.37,1.22,P＞0.05). Conclusions The development of temporal and spatial frequency characteristics of the rat LGN receptive field may be related to its functional visual pathway.

Background The afferent signals of proprioceptor in extraocular muscles play an important role in controlling eye position and conjugate movement. Palisade ending in the extraocular muscles is the main source of proprioceptive information, and its abnormalities in structure and function may be associated with the occurrence of nystagmus. Objective This study was to observe the changes of palisade ending in the extraocular muscles of patients with congenital nystagmus ( CN) and discuss the probable mechanism. Methods Modified Kestenbaum procedure was performed on 10 patients with CN, and the extraocular muscle samples were collected during the operation. Normal extraocular muscle samples were obtained from the enucleated eyeballs after ocular wound. The ultrathin sections of extraocular muscles were prepared and double-staining by uranyl acetate and lead citrate. The morphological changes of the palisade ending of extraocular muscles were examined under the transmission electron microscopy. Written informed consent was obtained from each subject before surgery. Results The ultrastructure of palisade ending in the extraocular muscle of CN subjects showed the different degrees of alterations. The mild changes included the collapse and disconnection of external capsules and the nonhomogeneous electron-dense substracts. The degeneration and dissociation of myelin in nerve endings, swelling and vacuolation of mitochondria were also exhibited. Myeloid body was found in axon. In the severe patients,the necrosis of Schwann' s cells,dissolve of axon and disappear of capsules were seen. Conclusion The palisade ending of extraocular muscle in the patients with CN are obviously abnormal in comparison with normal one. These alterations are probably associated with the etiology and pathogenesis of CN.

Firstly discovered in 1980s,human immunodeficiency virus (HIV) continues to affect more and more people.However,there is no effective drug available for the therapy of HIV infection.Betulinic acid existing in various medicinal herbs and fruits exhibits multiple biological effects,especially its outstanding anti-HIV activity,which has drawn the attentions of many pharmacists.Among the derivatives of betulinic acid,some compounds exhibited inhibitory activities at the nanomolar concentration,and have entered phase Ⅱ clinical trials.This paper summarizes the current investigations on the anti-HIV activity of betulinic acid analogues,and provides valuable data for subsequent researches.

Diabetic peripheral neuropathy (DPN), one of the most common chronic complications of diabetes, is characterized by allodynia, hyperalgesia and spontaneous pain. Chinese epidemiological studies have shown that at least 25% diabetic patients suffered from painful DPN, which compromises patients' daily functioning and becomes a major health care problem. Although the pathogenesis of painful DPN is not fully understood and current treatment options are very limited, research in the field has advanced our understanding on the mechanism of painful DPN in the past Decade of Pain Research and Control. This review will mainly focus on evaluation of current diabetic animal models, possible molecular pathways and available therapies, with an emphasis on roles of purinergic receptor and its signaling transduction pathways. Common therapies address one or two DPN symptoms, while others offer wider symptom control, presumably by targeting pathophysiological mechanisms of DPN. Purinergic receptor signaling transduction pathways might become potential targets for treatment for painful DPN.
Animals ; Diabetes Mellitus ; physiopathology ; Diabetic Neuropathies ; physiopathology ; Humans ; Hyperalgesia ; physiopathology ; Pain ; physiopathology ; Receptors, Purinergic P2X ; physiology

In the present study, a risperidone loaded microsphere/sucrose acetate isobutyrate (SAIB) in situ forming complex depot was designed to reduce the burst release of SAIB in situ forming depot and to continuously release risperidone for a long-term period without lagime. The model drug risperidone (Ris) was first encapsulated into microspheres and then the Ris-microspheres were embedded into SAIB depot to reduce the amount of dissolved drug in the depot. The effects of different types of microsphere matrix, including chitosan and poly(lactide-coglycolide) (PLGA), matrix/Ris ratios in microspheres and morphology of microspheres on the drug release behavior of complex depot were investigated. In comparison with the Ris-loaded SAIB depot (Ris-SAIB), the complex depot containing chitosan microspheres (in which chitosan/Ris = 1 : 1, w/w) (Ris-Cm-SAIB) decreased the burst release from 12.16% to 5.80%. However, increased drug release rate after 4 days was observed in Ris-Cm-SAIB, which was caused by the high penetration of the medium to Ris-Cm-SAIB due to the hydrophilie of chitosan. By encapsulation of risperidone in PLGA microspheres, most drugs can be prevented from dissolving in the depot and meanwhile the hydrophobic PLGA can reduce the media penetration effect on the depot. The complex depot containing PLGA microspheres (in which PLGA/ drug=4 : 2, w/w) (Ris-Pm-SAIB) showed a significant effectiveness on reducing the burst release both in vitro and in vivo whereby only 0.64% drug was released on the first day in vitro and a low AUC0-4d value [(105.2± 24.4) ng.mL-1.d] was detected over the first 4 days in vivo. In addition, drug release from Ris-Pm-SAIB can be modified by varying the morphology of microspheres. The porous PLGA microspheres could be prepared by adding medium chain triglyceride (MCT) in the organic phase which served as pore agents during the preparation of PLGA microspheres. The complex depot containing porous PLGA microspheres (which were prepared by co-encapsulation of 20% MCT) (Ris-PPm-SAIB) exhibited a slightly increased AUC0-4d of (194.6±15.8) ng.mL-1d and high plasma concentration levels from 4 to 78 days [Cs(4-78d)=(7.8±1.2) ng.mL-1]. The plasma concentration on 78 day C78d was (9.0 2.5) ng.mL-1 which was higher than that of Ris-Pm-SAIB [C78d= (1.6 ± 0.6) ng.mL-1]. In comparison with Ris-Pm-SAIB, the AUC4-78d of Ris-PPm-SAIB increased from (379.0±114.3) ng.mL-1.d to (465.0 ±149.2) ng.mL-1.d, indicating sufficient drug release from the Ris-PPm-SAIB. These results demonstrate that the risperidone loaded porous PLGA microsphere/SAIB in situ forming complex depot could not only efficiently reduce the burst release of SAIB depot both in vitro and in vivo, but also release the drug sufficiently in vivo, and be capable to continuously release the drug for 78 days.
Chitosan ; Drug Carriers ; Lactic Acid ; Microspheres ; Polyglycolic Acid ; Risperidone ; chemistry ; Sucrose ; analogs & derivatives ; Technology, Pharmaceutical