Objective To investigate the clinical features of glucose transporter 1 deficiency syndrome(GLUT1-DS) and summarize the characteristics of GLUT1-DS through reviewing related references.Methods The clinical data including manifestation,cerebrospinal fluid (CSF) glucose,electroencephalogram,MRI and gene mutation of a patient with GLUT1-DS was collected and the related literatures were reviewed.Results The patient was a 6 years old boy.The patient,whose seizures occurred at the age of 9 month-old and prolonged to 6 year-old,attacked before breakfast.Physical examination showed microcephaly with head circumference 47.5 cm.Laboratory tests showed that CSF glucose decreased (1.87 mmol/L) and CSF-serum ratio was 0.36.And meantime the MRI was normal and electroencephalogram showed general spike and slow wave complex paroxysm.Mutation of SLC2A1 gene,c.350_385del,was found in the patient.There were 219 cases with GLUT1-DS had been reported and the age of onset was 15.69 months.In 219 patients,159 cases (72％) suffered seizures,105 cases (47％) had motor abnormalities,61 cases (27％) suffered intellectual disability.The CSF glucose values were (1.92±0.31) mmol/L,CSF-serum ratio was 0.36±0.07.SLC2A1 gene mutations were detected in 183 patients(96％)in which missense mutation was the most mutation.Conclusion A wide range of phenotypes of GLUT1-DS include seizures,motor abnormalities,mental retardation.The diagnosis is confirmed when CSF glucose and CSF-serum ratio are continuously decreased which in the absence of meningitis.The SLC2A1 gene should be detected in suspicion of GLUTI-DS patients.Early diagnosis and treatment may improve the prognosis of those GLUTI-DS patients.

Mitochondrial dysfunction has been implicated in the aetiology of sporadic Parkinson's disease but its role in the disease mechanism remains unclear.To investigate the effect of synuclein on mitochondrial dysfunction induced by rotenone.The human dopaminergic SH-SY5Y cells were used as a cell model.The cells over-expressed the wild-type ?-synuclein were treated with complex I inhibitor rotenone.The cell viability,complex I activity,Mitochondrial swelling and O2-content were tested at different time point-1w,2w,4w after rotenone treated.CCK-8 test results showed that the cell viability of overexpressed ?-synuclein(SH-SY5Y-Syn)was much lower than the control group(SH-SY5Y-Ctr).After administrating with rotenone about 1w or 2w the cell viability of SH-SY5Y-Syn became higher than that of SH-SY5Y-Ctr.On the 4th week the results were contrary to the first 2 weeks.Similar results were got when test the mitochondrial function.In the first 2 weeks after roteoone administrating,the mitochondrial function of SH-SY5Y-Syn was better than that of SH-SY5Y-Ctr.This suggest that the ?-synuclein could protect the mitochondrial against the injury induced by rotenone in the early stage-1w,2w,while this effect disappeared in the final stage-4w.

Objective To explore the role of oxidative stress in the epithelial-tomesenchymal transition (EMT) of tubular epithelial cells and the protective effect of probucol in rat model with diabetic nephropathy (DN). Methods Thirty SD rats were randomly divided into normal control group, DN group, probucol treatment group (supplemented 1% probucol dietary). Twentyfour hours urinary protein excretion (UTP) was measured at the 3rd, the 8th and the 12th week respectively. The biochemical indicators including blood glucose (BG), lipids [triglyceride (TG), total cholesterol (TC)], low-density lipoprotein (LDL), serum creatinine (Scr), creatinine clearance rate (Ccr),kidney tissue malondialdehyde (MDA) level and glutathione peroxidase (GSH-Px) activity were assessed at the end of the 12th week in all groups. The renal pathological changes were evaluated by hematoxylin & eosin (HE) and Masson staining. The protein expression of specificity protein 1 (Sp1), α-smooth muscle actin (α-SMA) and E-cadherin was also detected and analyzed by immunohistochemistry and Western blotting. Results Compared with the normal control group,the BG, TC, LDL, Scr, 24 h UTP and MDA level of renal tissue increased significantly and the Ccr reduced in the rats of DN group (all P＜0.01). The pathological scores and the expression of Sp1 and α-SMA in renal tissue were higher in the DN animals than that in the other animals (all P＜0.01), the expression of E-cadherin downregulated significantly in the DN animals (P＜0.01). The MDA level of renal tissue was positively correlated to the expression of α-SMA and Sp1 protein in DN group (r=0.896, P＜0.01; r=0.862, P＜0.01, respectively), and negatively correlated to the expression of E-cadherin protein (r=-0.673, P＜0.01). In the diabetic animals treated with probucol, the Scr, 24 h UTP, pathological scores, MDA content,expression of Sp1 and α-SMA in renal tissue were lower than those in the diabetic animals (all P＜0.01). The Ccr and the expression of E-cadherin upregulated obviously (all P＜0.01). Conclusion Oxidative stress plays an important role in the EMT process of tubular epithelial cells. Probucol can slow down renal disease progression in DN rats through anti-oxidant, downregulating the expression of Sp1 protein and inhibiting the renal tubular EMT.

Objective To determine the effect of 2 transforming growth factor β1 (TGF-β1) short hairpin RNA (shRNA) expression plasmids (pcDU6-A1-A2 and pcDU6-B1-B2) on proliferation, TGF-β1, connective tissue growth factor (CTGF), and fibronectin (FN) expression induced by human serum albumin (HAS) in HK2 cells. Methods A vector plasmid containing the TGF-β1 shRNA was generated. An HK2 cell line was used in the study. The 2 TGF-β1 shRNA expression plasmids were transfected into cultured HK2 cells by lipofectamine 2000. Cellular proliferation was assessed by tetrazolium salt colorimetry. The semi-quantitative reverse transcriptive PCR was performed to detect the expression of TGF-β1,CTGF, and FN mRNA. Levels of TGF-β1 and FN protein were measured with a sandwich enzyme-linked immunosorbent assay. Results After treating with 5 g/L HAS for 24 hours in HK2 cells, cellular proliferating capacity increased significantly (P＜0.05). The expression levels of TGF-β1, CTGF, and FN mRNA were upregulated in HK2 cells stimulated by 5 g/L HAS, and levels of TGF-β1 and FN protein in the culture supernatant increased (P＜0.05). The introduction of pcDU6-A1-A2 and pcDU6-B1-B2 resulted in significant reduction of cellular proliferation activity, and the expression levels of TGF-β1, CTGF, and FN mRNA were downregulated (P＜0.05). Levels of TGF-β1 and FN protein in the culture supernatant decreased (P＜0.05) after 12 or 24 hours of TGF-β1 shRNA transfection into HK2 cells There was no significant difference in the expression levels of TGF-β1, CTGF, and FN mRNA between the 2 pcDU6 vector plasmid mediated TGF-β1 shRNA groups (P＞0.05). Conclusion pcDU6 vector plasmid mediated TGF-β1 shRNAs could obviously inhibit the expression levels of TGF-β1, CTGF, FN and cellular proliferation stimulated by HAS in HK2 cells, which may be related to the mediation of TGF-β1.

BACKGROUND: It is considered traditionally that epilepsy is a kind of complicated nervous conduct disorder caused by abnormally excited neuron in different area in brain. While the research on the function of astrocyte in epileptic attack is very rare.OBJECTIVE: To study the reaction of neuron and astrocyte in medullary visceral zone after epilepsy induced by pentetrazole in rats.DESIGN: A randomized controlled experimental research.SETTING and MATERIALS: The experiment was done in the Neurosurgery Laboratory of Zhujiang Hospital Affiliated to Southern Medical University and Neuroscience Institute of Fourth Military Medical University of Chinese PLA. Fourteen healthy adult SD rats, weighing 180 - 220 g, clean grade, were provided by the Experimental Animal Center of Fourth Military Medical University of Chinese PLA.INTERVENTIONS: Distribution of neuron and astrocyte in MVZ 1 hour after epileptic attack was shown by laserconfocal microscopic technique combined with triplication immunofluorescence histochemistry of anti-Fos protein, anti-tyrosine hydroxylase(TH) and anti-glial fibrillary acidic protein(GFAP).MAIN OUTCOME MEASURES: Observation of distribution of positive cells of Fos, GFAP and TH in MVZ and relationship between GFAP positive astrocyte and neuron.RESULTS: Fos positive neurons and GFAP positive astrocytes in MVZ increased significantly. Triplication immunofluorescence histochemistry showed reaction neuron(Fos positive) closely related with reaction astrocyte(GFAP positive) . Three kinds of N-ASC compounds with different labels were found, which were TH +/Fos +/GFAP + three labeled compound, TH + /GFAP +/Fos- and Fos+/GFAP +/TH- two labeled compound.CONCLUSION: Neuron and astrocyte in MVZ reacted strongly when epilepsy attacks. N-ASC as a functional unit may regulate onset of epilepsy.

Objective To determine the effect of smad anchor for receptor activation (SARA) on renal tubular epithelial to mesenchymal transtion (EMT) induced by high glucose and to investigate the associated mechanism.Methods HK-2 cells were exposed to high glucose (30 mmol/L).HK-2 cells were transfected with the plasmids of wild-type SARA [SARA (WT)] or SARA mutant [SARA with SBD deletion,called SARA (dSBD)] and then was stimulated by high glucose.The gene expression was assayed by real-time PCR and the protein expression was detected by Western blotting.Results During the process of high glucose-induced EMT of HK-2 cells,the gene and protein expression of SARA were down-regulated.The expression of TGF-β1 and Smad3 increased after stimulation of high glucose in HK-2.However,the Smad2 mRNA expression increased while its protein expression was down-regulated in a time-dependent manner.Smad2 and Smad3 were activated by high glucose stimulation and Smad3 kept activation for longer time than Smad2.Compared with high glucose group,over-expression of SARA by transfection of SARA (WT) up-regulated the expression of zona occludens(ZO)1 and down-regulated the expression of vimentin (P＜0.05).However,SARA (dSBD) had no such effects on above expressions.The Smad2 protein expression increased along with the over-expression of SARA.Meanwhile,over-expression of SARA prolonged the activation time of Smad2 and shortened the activation time of Smad3.Conclusions TGF-β1 signaling is activated and SARA expression is down-regulated during the process of high glucose-induced EMT in HK-2 cells.Over-expression of SARA can inhibit the EMT via increase of Smad2 protein expression and longer activation time of Smad2.

Objective To obtain differentiated osteoblast-specific inactivation of fgfr1 mice Methods To obtain fgfr1△/+/OC-CreTG/+ mice,fgfr1flox/flox mice obtained from NIH were crossed with OC-Cre mice To obtain fgfr1△/△/OC-CreTG/+ mutant mice,fgfr1△/+/OC-CreTG/+ further crossed with themselves or fgfr1flox/flox mice After fgfr1△/△/OC-CreTG/+ crossed with fgfr1flox/flox mice,half of their offspring were mutant mice Results Differentiated osteoblast-specific fgfr1 knockout mice were obtained Conclusion fgfr1△/△/OC-CreTG/+ mice were obtained through proper crossing strategy,which provides a suitable platform for studying fgfr1 function in bone development and fracture healing

Objective Discussing the role of Yunnan baiyao in the newborn umbilical cord.Methods 120 cases newborns were selected and randomly divided into the control group of 60cases and the observation group of 60 cases according to birth order.The two kinds of newborns were both used single valve care,and were disinfected with 75%alcohol twice a day,Newborns of the observation group were dipped in Yunnan baiyao cream which was taken 75%alcohol to apply the root of umbilical and around after each nursing time.Observed the umbilical cord healing of two groups of newborns and take notes.Results In the observation group newborns,umbilical cord on the drying time [(3.1 ±1.1)d]and off time[(6.6 ±1.8)d]were better than the control group[(3.1 ±1.1)d,(8.0 ±3.2)d](t =7.8,2.95,all P <0.05);6 cases occurred redness and swelling around the belly button,2 cases occurred hemor-rhage,17 cases occurred secretions.which were better than that in control group(18 cases,6 cases,35 cases).The statistical analysis showed significant difference in the data of the two groups.Conclusion Yunnan baiyao can promote healing of umbilical cord,it should be popularize in clinical use.

OBJECTIVETo observe the therapeutic effect and safety of perineal, crissal and progenital pruritus treated with acupuncture according to differentiation.

METHODSSelf-control method was applied in these 32 cases. Changqiang (GV 1), Huiyin (CV 1), Qugu (CV 2), Sanyinjiao (SP 6) and Ashi points etc. were punctured as main points, and adjunct points were added according to differentiation: Taichong (LR 3) and Ququan (LR 8) etc. were added for wind and heat excess of liver meridian, Xuehai (SP 10) and Quchi (LI 11) etc. were added for blood deficiency and wind dryness. Itchiness, skin lesions sign scores and therapeutic effects were observed before and after treatment.

RESULTSThe total scores of itchiness before and after treatment were 6.06 +/- 1.46 and 2.19 +/- 1.71 respectively, and the total scores of skin lesions sign were 4.38 +/- 2.21 and 1.50 +/- 1.44, indicating that the scores and the total scores of itchiness and skin lesions sign reduced obviously after treatment (P < 0.05, P < 0.01); the cured and markedly effective rate was 73.4% (11/15) for wind and heat excess of liver meridian, and 70.6% (12/17) for blood deficiency and wind dryness, presenting similar therapeutic effect (P > 0.05). Hematoma or ecchymosis appeared in 2 cases, and disappeared spontaneously after 2-3 days, without obvious adverse reaction.

CONCLUSIONSimple perineal, crissal and progenital pruritus treated with acupuncture according to differentiation is effective, safe and applicable.

Acupuncture Therapy ; Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Pruritus Ani ; therapy ; Pruritus Vulvae ; therapy ; Treatment Outcome ; Young Adult

Both genetic and environmental factors are involved in the pathogenesis of Parkinsonos disease (PD). Epidemiological studies showed that environmental factors shared with the common mechanisms of resulting in alpha-synuclein aggregation by inhibiting complex I of mitochondria and leading to oxidative stress. To investigate the relationship between alpha-synuclein and oxidative stress, we used human dopaminergic SH-SY5Y cells transfected with alpha-synuclein-enhanced green fluorescent protein (EGFP). alpha-synuclein gene expression was determined by immunocytochemistry and real-time quantitative PCR. Both SH-SY5Y and alpha-synuclein overexpressed SH-SY5Y (SH-SY5Y/Syn) cells were treated with various concentrations of rotenone for different time. Cell viability and oxidative stress were detected by MTT assay and DCF assay. Superoxide dismutase (SOD) activity was assessed with xanthine peroxidase method. Cell apoptosis was detected with flow cytometry. Results showed that alpha-synuclein gene was constantly overexpressed in SH-SY5Y/Syn cells. After treatment with rotenone, both cell viability and complex I activity in these cells were reduced in a concentration-dependent manner. Oxidative stress was also found in these cells. Compared with SH-SY5Y cells, SOD activity in SH-SY5Y/Syn cells was increased distinctly (P<0.05) and alpha-synuclein significantly attenuated rotenone-induced cell apoptosis. These results suggest that the alpha-synuclein overexpression in SH-SY5Y cells has a tendency to partially resist oxidative stress induced by rotenone and this response may assist cell survival.
Apoptosis ; drug effects ; Cell Line ; Cell Survival ; drug effects ; Cytoprotection ; Dose-Response Relationship, Drug ; Electron Transport Complex I ; metabolism ; Humans ; Oxidative Stress ; Rotenone ; toxicity ; Superoxide Dismutase ; metabolism ; Superoxides ; metabolism ; alpha-Synuclein ; genetics ; physiology