Objective To identify certain isoforms involved in the onset of retinal ischemic preconditioning (IPC), the effects of ischemic pretreatment (IP) were observed on level of conventional protein kinase C (cPKC) ?,?Ⅰ、?Ⅱand ? isoform-specific membrane translocation and protein expression in retina of rats. Methods Retinal IP was produced by intra-ocular pressure (IOP) elevation for 5 minutes in anesthetized Wistar rats. Sham operation was similar to IP except the pressure elevation. 10, 20, or 40 minutes and 1, 12, 24, 72 or 168 hours after the procedure, cPKC isoform-specific membrane translocation and protein expression were analyzed using Western-blot. Results cPKC? protein expression significantly increased from 12 h to 168 h. A peak reached at 72 h after IP. cPKC? membrane translocation enhanced during 20 min to 1 hours with a peak at 40 min. cPKC? protein expressionlevels significantly increased from 12 hours to 72 hours. A peak was found at 24 hours after IP. However, there were no significant changes in both membrane translocation of cPKC?, ?Ⅰ、?Ⅱand protein expression of cPKC?Ⅰ, ?Ⅱ in retina of rats following IP.Conclusion The enhanced cPKC? membrane translocation, the increased cPKC? and ? protein expressions might be involved in the onset and sustain of retinal IPC in rats respectively.

Objective To investigate low density lipoprotein receptor (LDLR)gene mutation in familial hypercholesterolemia (FH) patients. Methods The proband was given clinical diagnosis of homozygous FH based on marked features and blood lipid tests results. After apoB100R3500Q mutation was excluded, the promoter region and all of the 18 exons of LDLR gene were amplified by touch-downpolymerase chain reaction (PCR). The PCR products were analyzed by single-strand conformationalpolymorphism (SSCP). The PCR products with abnormal single strands were sequenced directly. Thesecondary structures of the mutational and wild type proteins were analyzed and compared byANTHEPROT5.0, and then the tertiary structures of the mutant and wild type LDLR were predicted atSWISS MODEL homepage online. Results A homozygous mutation A606T at exon 13 of the patients wasfound by SSCP and confirmed by DNA sequencing. GOR Ⅰ method in ANTHEPROT5.0 indicates that therandom coils and turns would replace some helixes at the mutation site. The online prediction from theSWISS MODEL homepage indicates the backbone structure of the mutant LDLR has no difference from thewild type one. Conclusion The results suggest the A606T mutation of LDLR gene is the cause of the FH inthis pedigree.

OBJECTIVETo screen the mutations of the low density lipoprotein receptor (LDLR) gene in a familial hypercholesterolemia (FH) family, and analyze the LDL-uptaking function of LDLR on lymphocytes of patients.

METHODSGenomic DNA was extracted from four affected members in a Chinese FH family. The presence of apoB100 gene R3500Q mutation which results in familial defective apolipoprotein B100 (FDB) was excluded first. Fragments of the LDLR gene were amplified by touch-down polymerase chain reaction (Touch-down PCR) and analyzed by single-strand conformational polymorphism (SSCP). The suspect fragments of the LDLR gene were cloned and sequenced. Furthermore, the lymphocytes bounded with fluorescent-labeled LDL (DiI-LDL) were measured by fluorescence flow cytometry.

RESULTSA nonsense mutation was identified in exon 10 of LDLR gene. This mutation gave rise to a premature stop codon (W462X), resulting in the absence of most of the LDLR domains. It was detected in all the affected members of the FH family. The ratios of functional LDLR in lymphocytes from patients and normal controls were 63.7% and 77.3% respectively. As a result, the activity of the functional LDLR in patients was just 82.4% of that in the normal controls.

CONCLUSIONIt is possible that the W462X mutation of LDLR gene is the main cause for the disease in this family.

Adult ; Apolipoprotein B-100 ; genetics ; Base Sequence ; Case-Control Studies ; DNA Mutational Analysis ; Deoxyribonuclease I ; metabolism ; Exons ; genetics ; Female ; Flow Cytometry ; Humans ; Hyperlipoproteinemia Type II ; genetics ; metabolism ; pathology ; Lipoproteins, LDL ; metabolism ; Lymphocytes ; metabolism ; Male ; Middle Aged ; Mutation ; Pedigree ; Receptors, LDL ; genetics ; metabolism

Objective: To investigate the diagnosis and clinical treatment of maxillofacial connective tissue hyperplastic trichoepithelioma. Methods: The clinical data of two cases of maxillofacial connective tissue hyperplastic trichoepithelioma were summarized and analyzed along with the literature Results : Two cases of maxillofacial connective tissue hyperplastic trichoepithelioma were male, aged 21 and 30 years. The clinical manifestations were painless pale brown and pale white plaques in the maxillofacial region. The lesion was tough and clear, with no ulcers in the middle depression. The course was 10-16 months, with 1-3 months before medical treatment, and the tumor had a significant history of enlargement. After surgery, the skin was cut 3 mm along the outer circumference of the tumor, and local tissue defects were repaired by the adjacent flap. The pathological report showed that the tumor cells were located in the dermis, and were striped, trabecular or nested. The tiny sac contained fibrous connective tissue proliferation. The tumor cells were amorphous without obvious nuclear division. Immunohistochemical analysis reported bcl-2(-), CK7(-), CK19(-), CD34(+), P63(+), CK56(+), and Ki67(±). The pathological diagnosis was connective tissue proliferative hair epithelial tumor. The patient was followed up for 24 months. There was no recurrence of the tumor, no obvious scarring, and no deformity or dysfunction of the maxillofacial region. Conclusion Pathological and immunohistochemical examination is the basis for the differential diagnosis of maxillofacial connective tissue hyperplastic trichoepithelioma, and surgical removal of tumors is an effective treatment.