AIM: To express the DT389-hbFGF (389 amino acid residues of the N-terminus of diphtheria toxin (human basic fibroblast growth factor) fusion protein for potential targeting therapy towards posterior capsule opacification (PCO) after cataract surgery.METHODS: The DNA of inactivated diphtheria bacillus and RNA of 12-week fetal brain cortex were extracted, respectively. The fragments of truncated diphtheria toxin (containing 389 amino acids of N-terminus, DT389) )and full-length human basic fibroblast growth factor(hbFGF) sequence (encoding 18kDa protein) were amplified by PCR. The two fragments were inserted into pGEX-4T-1 prokaryotic expression vector to obtain pGEX-DT389-hbFGF prokaryotic expression plasmid. After sequence analysis, the expressing plasmid was transformed into Escherichia Coli BL21 strain and expression was induced under IPTG. The expressed fusion protein was purified and identified.RESULTS: The gene fragments encoding DT389 and hbFGF were amplified and their gene sequences were confirmed. Hybrid gene expression plasmid pGEX-DT389 (hbFGF was constructed. The fusion protein DT389-hbFGF was expressed and purified.CONCLUSIONS: The successful cloning and expression of DT389-hbFGF immunotoxin provides a foundation for targeting therapy towards posterior capsule opacification.

Objective To evaluate the relationship between damages of visual field and retinal nerve fibre layer(RNFL)thickness in primary open angle glaucoma(POAG)with high myopia.Design Retrospective,case-controlled study.Participants POAG group with high myopia(21 eyes of 17 cases),POAG group with non-high myopia(17 eyes of 16 cases),high myopia group without POAG(25 eyes of 20 cases)and normal control group(19 eyes of 17 cases).Methods The static central visual fields were tested with Humphrey 750 automated perimetry and thickness of RNFL was measured with optical coherent tomography(OCT).Main Outcome Measures Mean deviation(MD),pattern standard deviation(PSD)and mean sensitivity at upper,lower,nasal and temporal quadrants in total deviation probability plots.Thickness of RNFL at upper,lower,nasal and temporal quadrants.Results There were more obvious general depression of sensitivity in total deviation probability plots of the early POAG with high myopia than those of POAG without high myopia,and the early visual field defects of glaucoma in pattern deviation probability plots of this group.MD of POAG with high myopia was more than those of others(P＜0.05).The differences of MD,PSD and mean sensitivity between POAG with high myopia and others were significant(P＜0.05).Mean sensitivities in each quadrant of POAG without high myopia were similar to those of high myopia(P＞0.05).The thickness of RNFL of POAG with high myopia was thinner than those of others and the thickness of RNFL of normality was thicker than that of others.The correlationship between mean sensitivity and the thickness of RNFL in each quadrant was significant(P＜0.05).Conclusion Pattern deviation probability plots is an important favor for judgement of the visual field changes in POAG with high myopia.The relationship between RNFL thickness with OCT and visual field damage may provide clinically relevant information in diagnosis of POAG with high myopia

Objective To investigate the correlation of the vascular endothelial growth factor (VEGF) gene variations in the promoter region with the sporadic Alzheimer' s disease (SAD) in Chinese Han population for better understanding the mechanism of SAD. MethodsThe polymorphisms of 279 SAD Chinese Han patients from Northern China were analyzed by comparing with those from 317 healthy individuals using the method of polymeraee chain reaction-restriction fragment length polymorphism ( PCR-RFLP) or direct sequencing.The commercial statistics package SPSS 11.5 was used to compare the distribution of the allele and the genotype, and to analyze their correlations with SAD. ResultsThree polymorphism sites were found for the VEGF promoters in the Chinese Han sample group including -2578C/A,- 2549I/D and- 1154G/A.- 2549I/D and- 2578C/A exhibiting strong linkage disequilibrium. Individuals with the A allele at position -2578 had an insertion of 18 nucleotides at -2459I/D, whereas CC homozygotes did not contain th es were found between the SAD patients and the controls in the 3 VEGF polymorphisms. After adjusting the data for gender, age and the ApoE ε4 allele using Logistic regression, the - 1154G/G genotype of the VEGF promoter might increase the risk of SAD in Chinese Han population.Among the subgroup without the ApoE ε4 allele, -2549D/-1154G haplotype might increase the risk for SAD (OR = 1.325, 95％ CI 1.023--1.716, P=0.033). ConclusionsThree polymorphism sites ( -2578C/A, -254911D, and -1154G/A) are found in the VEGF promoter regions in Chinese Han population. The-1154G/G genotype of the VEGF promote appears to increase the risk of SAD in Chinese Han population.In the absence of ApoE ε4, the -2549D/-1154G haplotype of the VEGF promoter appears to affect the risk for SAD.

Reversible cerebral vasoconstriction syndrome (RCVS) is characterized by thunderclap headache and reversible cerebral vasoconstriction, with other neurologic signs and symptoms. To the best of our knowledge, there were only a few cases of RCVS presenting both as both convexity subarachnoid hemorrhage (cSAH) and posterior reversible encephalopathy syndrome (PRES). Herein, we report a case of a 32-year-old woman with RCVS who presented with recurrent thunderclap headaches that occurred 50 days after delivery, with cSAH and PRES on magnetic resonance imaging (MRI). She had significant clinical and radiological recovery on 3 months’ follow-up. The clinical coexistence of cSAH and PRES in our case with RCVS is quite rare. This case illustrates the importance of awareness of the diagnosis of RCVS among clinicians even when initial brain and vascular imaging are normal. Early diagnosis and treatment are crucial for better prognosis.

Orally disintegrating tablets (ODT), a kind of new solid tablet that rapidly disintegrates to work in the mouth, has became the hot form of new drug research in recent years with many advantages, such as the convenient taking, a widely applicable people, fast acting, high bioavailability, good compliance, and so on. ODT has been widely used in chemical medicines, while the application of it in traditional Chinese medicines (TCMs) is still in the stage of development The development of TCMs ODT provides a new direction for the research of Chinese medicine new dosage, accelerates the pace of connecting to the world and modernization of Chinese medicine. This dosage has a broad market prospect, and its quality control and assessment standards, taste, the disintegration time in vitro and evaluation method are the key factors that affect the industrialization, standardization of Chinese medicine ODT. Therefore, this paper reviewed the characteristics, preparation, taste masking technology and quality evaluation with new technology of ODT. Meantime, numerous application examples of ODT used in traditional Chinese medicine were described. We expect to provide the reference and utilization for the development of traditional Chinese medicine orally disinteeratine tablets.
Administration, Oral ; Drug Compounding ; methods ; Humans ; Medicine, Chinese Traditional ; Solubility ; Tablets ; administration & dosage ; chemistry ; Taste

OBJECTIVE:To screen and identify endophytic fungi from Schisandra chinensis with antioxidant activity. METH-ODS:The tissue isolation skill was used to isolate endophytic fungi from roots,leaves,stems and fruits of S. chinensis. And anti-oxidant activity of endophytic fungi were screened by DPPH radical scavenging assay and hydroxyl radical scavenging assay. The to-tal DNA were extracted;the 18S rDNA ITS were amplified and sequenced with primer ITS1 and ITS4;the results of sequencing were analyzed comparatively based on homology to confirm the classification of active strains. RESULTS:23 strains were isolated from S. chinensis. GSR-12,isolated from roots of S. chinensis,had strong antioxidant activities. The scavenging rate on DPPH and the hydroxyl radical were 87.96% and 82.31% respectively. GSR-12 strain was identified as Clonostachys rosea by analyzed com-paratively. CONCLUSIONS:1 strain of C. rosea,isolated from roots of S. chinensis,has strong antioxidant activity.

The traditional Chinese medicine (Tripterygium wilfordiiHook.f.,TWH) has been clinically used to treat primary and secondary renal diseases and proteinuria for nearly 40 years.However,there is a rare literature about the effect of triptolide (the main active ingredient of TWH) on the expression of oxidative carbonyl protein (OCP) in diabetic nephropathy (DN).This study aimed to provide experimental evidence for triptolide treatment on DN through its effect on the expression of OCP,in order to investigate the effects of triptolide on the expression of OCP in rats with DN.Sixty SD rats were randomly divided into five groups:control group,high-dose triptolide (Th) group,low-dose triptolide (T1) group,DN model group,and positive control (benazepril) group.The DN model was established using streptozotocin.Urinary protein excretion,fasting blood glucose (FBG),superoxide dismutase (SOD) in renal homogenate,malondialdehyde (MDA) in renal homogenate and renal nitrotyrosine by immunohistochemistry,and the expression of OCP by oxyblotimmune blotting were detected.In the DN model group,rat urinary protein excretion and renal MDA were significantly increased,while renal SOD significantly decreased and nitrotyrosine expression was obviously upregulated in the kidney.After triptolide treatment,24-h urinary protein excretion (61.96±19.00 vs.18.32±4.78 mg/day,P＜0.001),renal MDA (8.09±0.79 vs.5.45±0.68 nmol/L,P＜0.001),and nitrotyrosine expression were decreased.Furthermore,renal OCP significantly decreased,while renal SOD (82.50±19.10 vs.124.00±20.52 U/L,P＜0.001) was elevated.This study revealed that triptolide can down-regulate the expression of OCP in the renal cortex of DN rats.

Background Retinal development continues during the early postnatal period in mammals.Correct arrangement of layers and precise location of various cells in the retina are vital for forming normal visual function during critical period plasticity.Spectral-domain optical coherence tomography(SD-OCT)provides highquality in vivo retinal imaging and the possibility to measure retinal thickness longitudinally. Objective The present study was to investigate the changes of retinal thickness during critical period plasticity in rats. Methods In vivo consecutive scanning of retinal image was performed in 10 SPF Sprague-Dawley rats at postnatal day 14(P14),P18,P21,P24 and P42 with SD-OCT,and retinal histopathological examination was used to detect retinal morphologic changes at the same postnatal ages in 20 matched rats.The whole retinal thickness,the thickness from inner limiting membrane(ILM)to inner plexiform layer(IPL),the thickness of inner nuclear layer(INL)and the thickness from outer nuclear layer(ONL)to retinal pigment epithelium(RPE)were measured using Cirrus HD-OCT system and HMIAS-2000 Imaging System in retinal sections.The measurement parameters by Cirrus HD-OCT and those by hematoxylin-eosin staining were compared.The use of animals followed the Statement of National Institute of Health (USA). Results In vivo high-resolution images of rat retinas with SD-OCT compared well with histology,which enabled quantitative comparison of the SD-OCT and histological data during critical period plasticity in rats.From P14 to P42,the retinal thickness gradually decreased with the increase of rat ages(F=15.425,P=0.000),and so were the thickness from ILM to IPL,the thickness of INL and the thickness from ONL to RPE(F=3.973,P=0.007;F=17.529,P=0.000;F=7.038,P=0.000).The retinal thickness,thickness of INL.thickness from ONL to RPE measured by Cirrus HD-OCT were significantly correlated with those measured by retinal sections among P14,P18,P21,P24 and P42 rats(r=0.794,P=0.000;r=0.784,P=0.000;r=0.681,P=0.000). Conclusion SD-OCT is a demonstratably valuable technology to study the structure of retinas in rats.The retinal thickness is shown to reduce in thickness throughout the development of the retina during critical period plasticity due to the decrease in thickness of INL and the distance from the ONL to RPE,as illustrated by OCT scanning.

AIM:To observe the structural basis of ocular motility abnormalities in patients with congenital fibrosis of the extraocular muscles type Ⅰ ( CFEOM Ⅰ) due to missense mutations in the developmental kinesin KIF21A using high - resolution magnetic resonance imaging ( MRI) .
METHODS: Totally 11 affected individuals reported KIF21A mutations were correlated with MRI studies demonstrating extraocular muscles ( EOMs ) size, location, contractility, and innervation. EOMs and the motor nerve in the orbits were imaged with T1 weighted in a triplanar scan using a dual-phased coils with 2. 0mm thick. Motor nerves were imaged at the brainstem using head coils and 3D-FIESTA with 0. 6-mm thick.
RESULTS: Patients with CFEOM Ⅰ exhibited different degrees of hypoplasia of oculomotor nerve, the abducens nerve and the trochlear nerve were also affected, of which 8 cases of orbital section could see the signal of abnormal nerve dominated by oculomotor nerve to lateral rectus. The both sides of six EOMS in all patients exhibited variable atrophy and abnormal bright internal signal on T1 imaging, particularly severe for the superior rectus and levator muscles.
CONCLUSION: High - resolution MRI can directly demonstrate pathology of motor nerves,affected EOMs, and ‘Pulley' hypoplasia caused by CFEOM Ⅰ due to mutations in KIF21A,and these findings suggest that the neuronal hypoplasia is the etiological factor of CFEOM.

OBJECTIVE: To observe the effects of intermedin preconditioning on hypoxic injury in rat's cardiac myocytes and to provide the hypothetical mechanism of sudden cardiac death in the field of forensic pathology. METHODS: The H9c2 cultured rat cardiac myocytes were randomly divided into control group, hypoxia group and IMD group. The myocardial cell viability, cellular ultrastructure, intracellular calcium concentration and apoptosis rate were determined by MTT assay, transmission electron microscopy, laser scanning confocal microscope and flow cytometry, respectively. RESULTS: Compared with the control group, cell viability obviously decreased with inner ultrastructure injury in the hypoxia group (P<0.05), while cell viability significantly increased in the IMD group by reducing the hypoxia injury of cardiac myocytes (P<0.05). Compared with the control group, [Ca2+]i (fluorescence intensity) and apoptosis rate significantly increased in the hypoxia group, but decreased in the IMD group (P<0.05). CONCLUSION IMD increases the cell survival rate and decreases the cell apoptosis inhibited by intracellular calcium overload from hypoxia. This finding may reveal the mechanism of protective effects of myocardial hypoxia, and provide a scientific basis for the identification sudden cardiac death.
Animals ; Apoptosis ; Calcium ; Cell Hypoxia ; Cell Survival ; Hypoxia ; Myocardial Ischemia ; Myocardium/cytology* ; Myocytes, Cardiac/physiology* ; Rats ; Rats, Sprague-Dawley