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.

Objective To investigate the effects of different durations of methamphetamine abuse on spatial cognitive processing.Methods The mental rotation task was used to evaluate the spatial processing function of 35 methamphetamine abusers and 30 healthy subjects.The methamphetamine abusers are divided into 2 groups according to their abuse durations,group 1 contains 16 abusers with an average duration of 1 year and group 2 contains 19 abusers with an average duration of 3 years.Participants were asked to judge the right hand/food or the left hand/food of the experimental stimuli at difference angles.The reaction time (RT) and the accurate rate (AR) of the mental rotation task were recorded.Results Compared with the control group,RT of group 1 methamphetamine abusers was longer at the angle of 0°((1469±318)ms) and 180°((1718±412)ms) (P＜0.05),RT of group 2 was longer at the angle of 0°((1466±243) ms),60°((1497±294) ms) and 180°((1708±288) ms) (P ＜0.05).And the AR of methamphetamine abusers was higher at every angle (P＜0.05).Conclusion Long-term dependence on methamphetmine can damage the abuser's spatial processing function.

Objective To investigate whether conventional protein kinase C (cPKC ) βⅡ-interacting collapsin response mediator protein-2 (CRMP-2) provides neuroprotection against cerebral ischemic (I) injuries. Methods Male BALB/c mice were randomly divided into normoxic control (Nor) , HPC, Nor + Sham, HPC + Sham, Nor + I and HPC + I groups (n = 6 per group). Using our HPC and MCAO mouse models, we applied immunoprecipita-tion, two-dimensional electrophoresis and mass spectrometry to characterize cPKCβⅡ-interacting proteins and combined with SDS-PAGE and Western blot to quantitatively analyze CRMP-2 phosphorylation and degradation levels in the brain of mice after HPC and MCAO. Results The expression level of 10 cPKCβⅡ-interacting proteins changed obviously in cerebral cortex of HPC mice when compared with Nor group. One of these proteins, CRMP-2 protein level increased in particulate fraction and decreased in cytosolic fraction of cerebral cortex of HPC mice. CRMP-2 phosphorylation level in ischemic core (Ic) of cerebral cortex decreased significantly ( P < 0. 05 , n = 6) as compared with that of Nor + sham group, but CRMP-2 phosphorylation level in HPC +I group increased significantly as compared with that of Nor +I group ( P < 0. 05, n = 6). In ischemic cortex, CRMP-2 degradation (proteolysis) was observed as the appearance of 55 ku breakdown products (BDP). However, the CRMP-2 degradation level, BDPs products decreased significantly in penumbra ( P) of ischemic cortex from HPC +I group when we compared with that of Nor +I group (P < 0. 05, n = 6 ). Conclusion CRMP-2 is involved in attenuating the decrease of CRMP-2 phosphorylation in ischemic core and in inhibiting its degradation in penumbra of cerebral cortex of mice thereby to lessen the ischemic injuries.

Objective To investigate the risk factors related to progressive intracranial hemorrhage (PIH) in patients with acute traumatic brain injury (TBI) and analyze their clinical significance.Methods PIH was validated by comparing the initial and repeated CT scans. Data including gender,age, injury causes, Glasgow Coma Score (GCS) on admission, time interval from injury to the first CT scan, initial CT scan manifestations, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fg), thrombin time (TT), platelet (PLT) and D-dimer (D-D) in both groups were compared with Logistic regression analysis to observe the risk factors related to PIH. Results The study involved 498 patients with acute TBI, of which 139 (27.91%) patients suffered from PIH. There were 116 patients (83.45%) with PIH who received the initial CT scan within two hours post injury.There was statistical difference in aspects of age, GCS on admission, time interval from injury to the first CT scan, initial CT scan manifestations ( including fractures, subarachnoid hematoma, contusion and onset hematoma), PT, Fg and D-D values in both groups (P ＜0.01 ). Logistic regression analysis showed that CT scans (subarachnoid hemorrhage, brain contusion and primary hematoma) and plasma D-D values were predictors of PIH ( P ＜ 0.01 ). Conclusions For patients with the initial CT scan manifestations including subarachnoid hemorrhage, brain contusion, primary hematoma together with D-D value increase within two hours post injury, a continuous CT scan should be performed promptly to detect PIH early.

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

We expressed B cell epitopes of dengue virus envelope protein and NS1 protein in prokaryotic cells,and purified and evaluated for its serological activities.A recombinant multi-epitope chimeric gene named rE including eight B cell epitopes was connected by linker peptide (EAAAK)2 and cloned into prokaryotic expression vector pET-28a(+),and transformed into E.coli BL21(DE3) cells for expression under induction of IPTG.The expressed recombinant protein was purified with 6× His purification media,and identified by SDS-PAGE and Western blot,and its antigenicity was analyzed by using an indirect ELISA assay.The recombinant expression vector pET28a-rE was constructed and expressed in BL21 (DE3) successfully,but the recombinant proteins mainly appeared as inclusion bodies.The target protein was obtained with high purity through the purification of affinity chromatography.SDS-PAGE and Western blot analysis showed that the molecular weight of fusion protein was in the expected line.The established indirect ELISA has high accuracy.This recombinant peptide antigen expressed in E.coli has good potential for serum testing.

Objective To investigate the clinical effect of minimally invasive extraction of anterior tooth residual root after root separation.Methods A total of 400 patients receivinganterior tooth residual root extraction were collected in the clinic of oral and maxillofacial surgery department between January 2015 and December 2016.The patients were divided into a control group and a study group according to their sequence to see the doctor,with an odd for the study group and an even for the control group.In the study group,residual roots were separated mesiodistally by high speed turbine before using minimally invasive extraction tool;while in the control group residual roots were extracted only using minimally invasive extraction tool.The surgical duration,postoperative damage rate of the lip side plate,degree of pain and patient satisfaction in the two groups were analyzed.Results The surgical duration was shorter in the study group compared with the control group (P ＜ 0.05).The postoperative damage rate of the lip side plate and the degree of pain were lower,while patient satisfaction was higher in the study group than in the control group (P ＜ 0.05).Conclusions The postoperative damage rate of the lip side plate is significantly lower in minimally invasive extraction of anterior tooth residual root after root separation.Smaller trauma is conducive to the implant afterwards.Root separation in minimally invasive extraction of anterior tooth residual root is valuable for clinical application.

Objective:To explore the relationship between the changes of anorectal angle (ARA) under 3 physiological states and pelvic organ prolapse(POP) in postpartum women by transperineal ultrasound.Methods:The retrospective study enrolled 147 female in 6-8 weeks after delivery examined by pelvic floor ultrasound examinations in Fujian Medical University Second Affiliated Hospital from November 2019 to June 2021, who were divided into POP group and control group. Volume data of pelvic floor ultrasound examinations were obtained at rest, during contraction and during maximal Valsalva maneuver. The differences in the changes of ARA under 3 physiological states between the two groups were compared, and the correlation between the change state of ARA during maximal Valsalva maneuver and POP was analyzed.Results:Compared with ARA at rest, ARA decreased during contraction (χ 2=42.64, P<0.001) and increased during maximal Valsalva maneuver (χ 2=38.43, P<0.001). There was no difference of ARA between the POP group and control group in the 3 physiological states ( P>0.05). However, the risk of POP increased when ARA decreased during maximal Valsalva maneuver ( OR=2.690, 95% CI=1.074-6.739, P<0.05). Conclusions:The decrease of ARA during maximal Valsalva maneuver may increase the risk of POP, and the change of ARA during maximal Valsalva maneuver can be brought into the ultrasonic observation indicators of POP.

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.

Type 1 diabetes mellitus (T1DM)-induced cognitive dysfunction is common, but its underlying mechanisms are still poorly understood. In this study, we found that knockout of conventional protein kinase C (cPKC)γ significantly increased the phosphorylation of Tau at Ser214 and neurofibrillary tangles, but did not affect the activities of GSK-3β and PP2A in the hippocampal neurons of T1DM mice. cPKCγ deficiency significantly decreased the level of autophagy in the hippocampal neurons of T1DM mice. Activation of autophagy greatly alleviated the cognitive impairment induced by cPKCγ deficiency in T1DM mice. Moreover, cPKCγ deficiency reduced the AMPK phosphorylation levels and increased the phosphorylation levels of mTOR in vivo and in vitro. The high glucose-induced Tau phosphorylation at Ser214 was further increased by the autophagy inhibitor and was significantly decreased by an mTOR inhibitor. In conclusion, these results indicated that cPKCγ promotes autophagy through the AMPK/mTOR signaling pathway, thus reducing the level of phosphorylated Tau at Ser214 and neurofibrillary tangles.
AMP-Activated Protein Kinases/metabolism* ; Animals ; Autophagy ; Diabetes Mellitus, Type 1 ; Glucose ; Glycogen Synthase Kinase 3 beta/metabolism* ; Mice ; Phosphorylation ; Protein Kinase C/metabolism* ; TOR Serine-Threonine Kinases/metabolism* ; tau Proteins/metabolism*