Corneal dystrophy represents a group of inherited corneal diseases with progressive accumulation of deposits in different layers of cornea,resulting in loss of corneal transparency and visual impairment,or even blindness.Initial classification of corneal dystrophy was upon involved cornea layer,and it is insufficient for some multi-layer lesion.With the current progress in molecular genetics,researchers proposed a new classifying way based on genetic information of corneal dystrophy.A revised classification has been recommended by the International Committee for Classification of Corneal Dystrophies (IC3D).The clinical manifestation,histological and genetic basis of the disease are integrated in the classification system,so it is regarded as to be more scientific and reasonable.Recent years,our laboratory performed genetic screen on some Chinese pedigrees with corneal dystrophy,mainly focusing on the corneal dystrophy-associated genes such as human transforming growth factor beta induced (TGFBI) gene,which reveals the relationship of the different mutations on TGFBI gene with clinical phenotypes.Our studies further indicated that the corneal dystrophy classification method based on molecular level is a more scientific and practicable method.Some updated information on the clinical phenotypes and molecular aspects of corneal dystrophy were provided here aimed to offer the aid to the differential diagnosis and management of these diseases.

Advance on chemical constituents and pharmacological activities of Stellera plants have been conducted. The chemical constituents include terpenes, coumarins, flavonoids, lignans, volatile oils, and other compounds. Pharmacological studies showed that diterpenoids and biflavones showed strong activities, such as antitumor, anti-HIV, and immune regulations. This review hopes to provide a scientific basis for further research and explorations of the medicinal values of the genus.
Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Molecular Sequence Data ; Molecular Structure ; Thymelaeaceae ; chemistry ; classification

Objective To know the indication for patients with spinal cord injury during the course of intermittent urethral catheterization.Methods Divided 33 patients with spinal cord injury into the experimental group(18 cases)and control group(15 cases)randomly.The indication of beginning in the experiment group was less than 500 ml transfusion per day,without press ulcer,more than 150 ml bladder capacity.The indication in the control group was＞28 cm H2O pressure of bladder.Compared the effects between the two groups.Results The incidence rate of infection in the experiment group was lower than control group,all the indexes of uretharal catheterization were better in the experiment group than those of in the control group.Conclusions The indication of less than 500ml transfusion per day,without press ulcer,more than 150 ml bladder capacity are proper.

Background Human paired box gene 6 (PAX6)encodes a transcriptional regulator.It is essential for eye and brain morphogenesis.Mutation of PAX6 gene isresponsible for many congenital ocular malformations,such as aniridia.Aniridia is a autosomal dominant inheritance mode.Objective In this study,PAX6 gene mutation was analyzed in three Chinese families with aniridia through polymerase chain reaction (PCR) and sequencing.Methods The blood specimens were collected from 5 suffers and normal individuals of 3 aniridia families to extract DNA.The sequences of extron 4-13 were designed based on PAX6 gene.The primer was amplified by PCR and sequenced and compared with the known PAX6 gene sequence.This study complied with Declaration of Helsinki and approved by ethic committee of Sichuan University.Written informed consent was obtained from each individual before any medial examination.ResultsThere were 5 suffers in the 3 families.A heterozygous mutation (c.718 C＞T) in PAX6 gene was identified in 2 patients of family A.This mutation caused an amino acid substitution of arginine to termination codon at position 240 ( p.Arg240X) of PAX6 protein.No similar change in the normal families.No any the alteration of PAX6 gene was detected in family B whatever suffers and normal individuals.In family C,a deletion mutation of c.331 delG ( p.Val111 SerfsX13 ) in PAX6 gene was found.The deletion of one base caused frame shift mutation of PAX6 protein,and no such mutation was seen in other families.Conclusions Mutation of PAX6 gene appeares to be causative mutations of the disease in family A and C.

Background Posterior capsular opacification(PCO) is common complication after extrecapsular extract of cataract.Matrix metalloproteinases-3 (MMP-3) can degrade all the extracellular matrix except polyose.The gene therapy of PCO upon MMP-3 is the researching hot topic.Fibronectin ( FN ) is a degrade gelatin,so its expression can reflect the effect of MMP-3 on LECs indirectly. Objective The aim of this study was to construct MMP-3 eukaryotic recombination plasmid and transfect to lens epithelium cells(LECs) for the observation of MMP3 expression,and to explore the feasibility of gene therapy for after cataract. Methods Six fresh lenses were obtained from pigs.LECs were cultured using explant method.The eukaryotic expression vector pEGFP-N1-MMP-3 was reconstructed with MMP-3 and pEGFP-N1 plasmids.The accuracy of MMP-3 gene fragment was confirmed by double enzyme digestion and DNA sequencing analysis.After transfecting pEGFP-N1-MMP-3 into LECs of pig,the expression of MMP-3 protein in the cells was indirectly observed by green fluorescent protein.The expression of FN in LECs was detected using Western blot. Results The result of double enzyme digestion was consistent with the base number of pEGFP-N1 plasmids and target fragment.By enlacing the result of DNA sequencing analysis with software,the resemblance of the DNA sequence of MMP-3 from recombination plasmid pEGFP-N1-MMP-3 and that of homo MMP-3 was 99.6％,indicating that the target fragment was inserted to pEGFP-N1 plasmids successfully.Green fluorescence for GFP was seen in the LECs in pEGFP-N1-MMP-3 transfected group,but absent response for GFP was in empty vector group.Western blot revealed that the relative expression level of FN in LECs was 0.666±0.008 in pEGFP-N1-MMP-3 trasfected group and 0.326 ±0.071 in empty vector group,with a significant difference between these two groups(P=0.000). Conclusions Eukaryotic recombination plasmid pEGFP-N1-MMP-3 is successfully constructed,and MMP-3 can be expressed in LECs after transfected.These results lay a foundation for the further research of MMP-3 gene therapy for PCO.

OBJECTIVESTo construct the expression vector pGenesil-3-shRNA that can express the short hairpin RNAs (shRNA) silencing Kir6.2 gene and to study its influence on the proliferation and invasion of HepG2 cells.

METHODSTwo shRNA silencing Kir6.2 genes were transcript synthesized intracellularly by expressed templates of plasmid vector pSilence-3, and the target sequence of Kir6.2 gene was inserted into the upstream of the reporter gene in order to construct the recombinant plasmid vector pGenesil-3. Plasmids containing 2 different sequences of human Kir6.2 mRNA coding region were constructed and transfected into HepG2 cells by using lipofectamine 2000 methods. The experiment was divided into 4 groups: SK (normal), SK-HK (negative control), SK-K1 (transfected with the interfering sequence 1) and SK-K2 (transfected with the interfering sequence 2) groups. A selected single clone was cultured after screening by G418. The expression of Kir6.2 protein was detected by Western blot. MTT assay and Transwell system were used to observe the proliferation and invasion of HepG2 cells.

RESULTSThe recombinant expression plasmid pGenesil-3 was successfully constructed and underexpression of Kir6.2 gene in HepG2 cells was detected by Western blot. Underexpression of Kir6.2 gene significantly decreased the proliferation and invasion of the HepG2 cells.

CONCLUSIONshRNA can inhibit the expression of Kir6.2 gene in the HepG2 cells, and underexpression of Kir6.2 gene decreased the proliferation and invasion of the HepG2 cells.

Cell Proliferation ; Genetic Vectors ; Hep G2 Cells ; Humans ; Neoplasm Invasiveness ; Plasmids ; Potassium Channels, Inwardly Rectifying ; genetics ; RNA, Small Interfering ; genetics

Objective To investigate the effect of different doses valsartan on atherosclerosis in rabbits. Methods 53 white rabbits were randomly divided into control group,cholesterol group,and valsartan group of high-,middle-,and low-dose.The positive expression of pro-oncogene c-myc and c-fos mRNA was studied with in situ reverse transcription-PCR)ISRT-PCR).The histoppathological changes of aorta were observed.Results)1) Positive expression rate of pro-oncogene c-myc and c-fos mRNA was significantly higher in cholesterol than that in the qontrol group.The rate of positive expression was remarkably lower in the high-dose valsartan group than that in the cholesterol group)P

5.5mmol/L),persistent metabolic acidosis,or low cardiac output syndrome.Following data were collected in all patients:time to ni- tiation and duration of PD;time point of the recovery of urine output;baseline serum creatinine level(Cr0),rise of Cr(Crl),peak Cr (Cr2),descending Cr(Cr3),recovery of Cr(Cr4);and their corresponding postoperative time points.Results Of the 63 patients,58 (92.1%)required PD.Overall mortality rate was 33.3%(21/63).Patients undergone more complex surgery requiring longer aortic damping time;have higher Cr0,Cr2,Cr3 and longer period of the recovery of Cr and urine output(P6d)was associated with more complicated surgical procedure,higher Cr1 and Cr2,delayed recovery of Cr and urine output after surgery,longer period of low cardiac output syndrome,more dysfunctional organs,longer mechanical ventilation and ICU stay postoperatively(P

OBJECTIVETo investigate the toll-like receptors (TLR) profile of human epidermal keratinocytes.

METHODSWe cultured the immortalized human epidermal keratinocyte cell line HaCaT cells and normal human epidermal keratinocytes (NHEK) and separated epidermis with dispase from foreskins. TLR 1-10 mRNA expression was detected with reverse transcription polymerase chain reaction (RT-PCR). TLR 2 and 4 protein expressions on surface of HaCaT cells and NHEK were detected using flow cytometry.

RESULTSHaCaT cells, NHEK, and epidermis all expressed TLR 1-10 mRNA with different intensity. TLR 4 protein was detected on the surfaces of HaCaT cells and NHEK, while the expression of TLR 2 protein was few.

CONCLUSIONHuman epidermal keratinocytes constitutively express all TLR 1-10 mRNA, which may enable human keratinocytes to respond to a wide range of pathogenic micro-organisms.

Cell Line ; Cell Line, Tumor ; Epidermis ; cytology ; Flow Cytometry ; Humans ; Keratinocytes ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Toll-Like Receptors ; genetics ; metabolism

AIMTo investigate the effects of hepatocyte growth factor gene transfected MSCs transplantation on cardiac function and fibrosis in rats heart failure model induced by adriamycin.

METHODSMSCs were isolated from SD rats by density gradient centrifugation, purified, and transfected with Ad-hHGF. ELISA were used to detect the protein expression of hHGF in these MSCs. Forty SD rats underwent intraperitoneal injection with adriamycin to induce heart failure model. 8 healthy rats served as control, 24 survival rats were randomly divided into 3 groups (n = 8): Rats in Ad-hHGF transfected MSCs group were injected with Ad-hHGF transfected MSCs 2 weeks after the establishment of the model, rats in MSC group injected with suspension of MSCs only, and model group was injected with cold culture fluid. Heart function was evaluated by a physiological recorder 4 weeks after cell transplantation. Myocardial cell morphology and interstitial collagen were studied by electron microscope and were stained by Sirus red. TGF-beta1 was detected by immunohistochemical method.

RESULTS(1) MSCs could be transfected efficiently by Ad-hHGF, manifested by a higher level of expression in vitro, persisting 14 days at least. (2) Four weeks after the cells transplantion, cardiac necrosis in MSC-hHGF rats was improved when compared with those in the MSCs (P < 0.05) and Model group (P < 0.01). The heart function of the MSC-hHGF rats was greatly improved with an significant increase in LVSP and + dp/dt(max), although LVEDP still highter than that of normal rats. (3) MSC-Ad-hHGF decreased Myocardial collagen content and the level of TGFbeta1 compaired with MSCs transplanted rats (P < 0.01).

CONCLUSIONTransplantation of HGF gene transfected MSCs improved heart function, decreased myocardial collagen and the level of TGFbeta1.

Animals ; Doxorubicin ; Fibrosis ; metabolism ; prevention & control ; Heart Failure ; chemically induced ; physiopathology ; therapy ; Hepatocyte Growth Factor ; genetics ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; metabolism ; Myocardium ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transfection