Objective To identify mutations site and clinical characteristics of a familial hypercholesterolemia(FH) proband diagnosed clinically through DNA sequencing and family analysis in the proband and his family members of 3 generations.Methods Blood samples and clinical data of the kindred of total 29 from 3 generations members were collected.Proband had a physical examination electrocar-diogrom and vascular ultrasound.The proband and his family members took routine clinical exams,and genomic DNA was isolated.The promoter region and the 18 exons of low density liporotein receptor(LDLR) gene were screened by Touch down polymerase chain reaction -single strand conformation polymorphism(PCR-SSCP) and DNA sequencing.The result of sequencing were matched gene sequence published in the BLAST database.Results 1.Increased intima-media thickness and plaque were detected in the common carotid artery,right subclavian artery of the proband.Aortic valve regurgitation was found by echocardiography.2.No mutation R3500Q of ApoB100 was observed.3.Two heterozygous mutations in exon 10 and 13 of LDLR gene (W462X and A606T) were identified.The proband and 5 members of paternal relatives showed W462X heterozygosis mutation in exon 10 of LDLR gene which introduced the change from tryptophone to a new stop codon.The proband's mother and grandmother harboured A606T heterozygous mutation in exon 13 of LDLR gene due to a single base pair substitution of G for A in the codon for residue 1 879.Conclusions Disease causing mutations of proband are W462X and A606T compound heterozygosis mutation in exon 10 and 13 of LDLR gene inherited from mother and father.Proband shows homozyous phenotype though the genotype analysis indicates heterozygous mutations.

Objective To study the effect of selenium on peripheral and splentic T-cell subset, apoptosis of spleen cells in fluorosis chicken and its mechanism. Methods One hundred and eighty 8-day Hailanhe chicks were randomly divided into 3 groups(each 60): ①control group: 195 mg/kg fluoride and 0.08 mg/kg of selenium; ②fluorine group : 1000 mg/kg fluoride and 0.08 mg/kg of selenium ;③selenium antagonism group : 1000 mg/kg On 30~(th), 60~(th), 90~(th) day, peripheral and splentic CD4~+, CD8~+ T-cell subset analyses underwent flow cytometry and apoptosis of spleen cells were detected by TUNEL for study subjects. Results Compared with control group, the CD4~+ T-cell subset of peripheral in fluorine group was decreased obviously in 30,60,90 days[ (35.36± 4.27)% vs (24.29 ± 2.96)%, (47.65 ± 5.42)% vs (41.62 ± 3.96)%, (49.58 ± 3.98) % vs (42.35 ± 6.03 )%, P < 0.05 or < 0.01 ], CD4~+/CD8~+ ratio also was decreased obviously [ ( 1.701 ± 0.145 )% vs (1.393 ± 0.163)%,(2.712 ± 0.345)% vs (1.781 ± 0.201)%,(2.438 ± 0.356)% vs (1.973 ± 0.229)%, P< 0.05 or < 0.01]. Compared with fluorine group, the CD4~+ T-cell subset of peripheral in selenium antagonism group [ (29.40 ± 3.38)%, (45.40 ± 6.01 )%, (46.85 ± 5.25)%, P < 0.05 or < 0.01 ] was increased obviously in 30,60,90 days,CD4~+/CD8~+ ratio in 60,90 days[(2.004 ±0.314)%,(2.211±0.229)%,all P<0.01]also was increased obviously.Compared with control group,the CD4~+ T-cell subset of spleen cells in fluorine group was decreased obviously in 30,60,90 days[(47.33±5.35)% vs(41.91±4.83)%,(49.28±5.24)% vs(41.26 ±4.56)%,(34.31±4.15)%vs(29.33±2.89)%,all P<0.01],CD4~+/CD8~+ ratio also was decreased obviously[(1.927 ±0.244)% vs(1.525 ±0.265)%,(1.847±0.224)% vs(1.640±0.198)%.(1.265±0.174)% vs(0.878±0.092)%,P<0.05 or<0.01].Compared with fluorine group,the CD4~+ T-cell subset of spleen cells in selenium antagonism group in 60,90 days[(44.87±5.43)%,(32.62±3.37)%,all P<0.05]was increased obviously,CD4~+/CD8~+ ratio in 30,60, 90 days[(1.703 ±0.201)%,(1.772±0.215)%,(0.991±0.124)%,P<0.05 or<0.01]also was increased obviously. The apoptosis ratio of spleen cells in fluorine group in 30,60,90 days[(2.31±0.36)%,(2.76±0.22)%,(3.04± 0.29)%]was higher than that in control group[(1.14±0.21)%,(1.23±0.23)%,(1.29±0.20)%,P<0.01].The apoptosis ratio of spleen cells in selenium antagonism group in 60,90 days[(2.42 ±0.32)%,(2.73±0.39)%]was lower than that in fluorine group(P<0.05 or<0.01).Conclusion A certain concentration of selenium can antagonize the immunity inhibition of fluorine by decreasing apoptosis and improving the unbalance of T-cell subset.

Objective To analyze the awareness on health among high school students and its influencing factors in Guangdong. Methods Multi-stage sampling and questionnairs "2009 health awareness survey of the Chinese citizens" developed by our Department of Health, were used. Data were analyzed by multivariate multilevel model under MLwinN 2.19 software. Results The mean scores on knowledge and ideas, behaviors and related skills among 1606 high school students of Guangdong province, were 69.08 ± 14.81,60.05 ± 16.85 and 74.99 ± 21.17 respectively. Three items on health showed that they all related to each other and relations between grades (0.972,0.715 and 0.855) were greater than the individuals (0.565,0.426 and 0.438). Conclusion Factors as students from outside the Pearl River Delta region or from the rural areas, being male, at general secondary schools, at grade one, with poor academic performance and more pocket money etc., had lower levels on those related information of health.

AIMTo clone and analyse the coding cDNA sequence of alpha1, beta2 and gamma2 subunit of GABAA receptor in American king Pigeon.

METHODSWithdrew total RNA from the American king pigeon brain, reverse transcribing general primers to acquire a gene set cDNA. Designing specific primers of three subunit mRNA of the GABAA receptor, by RT-PCR respectively expanded the conservative gene of al subunit, beta2 subunit and gamma2 subunit of GABAA receptor, and carried on clone, plastid identification and the sequence measurese of three genes.

RESULTSThe experiment on sequence measures has succeeded that sequence analysis indicated that lengths of the conservative gene of alpha1 subunit, beta2 subunit and gamma2 subunit of GABAA receptor was respectively 899 bp, 597 bp and 563 bp, homology on reference sequence was respectively 94.99%, 94.64% and 96.28%.

CONCLUSIONHomology is high on the conservative gene of alpha1 subunit, beta2 subunit and gamma2 subunit of GABAA receptor of brain tissue of pigeon and chicken but there is a discriminating characteristic in different kinds of animals.

Animals ; Brain ; metabolism ; Cloning, Molecular ; Columbidae ; DNA, Complementary ; genetics ; RNA, Messenger ; genetics ; Receptors, GABA-A ; classification ; genetics ; Sequence Analysis, DNA

OBJECTIVENuclear factor-kappa B (NF-kappaB) is a critical transcription factor governing the expression of many cytokines that are involved in the pathogenesis of inflammatory diseases, such as asthma and rheumatoid arthritis. Melatonin (MT), a relatively safe and potent antioxidant which has shown efficacy in several chronic inflammatory models, may inhibit the expression of NF-kappaB and therefore might have a therapeutic use in asthma. This study aimed at observing the effect of MT on the expression of NF-kappaB and airway inflammation in a rat model of bronchial asthma.

METHODSTwenty-four male Sprague-Dawley (SD) rats weighing 120 g to 170 g were randomly divided into three experimental groups (8 in each): (1) Asthmatic group: Rats were immunized on day 1 by intraperitoneal injection of 100 mg ovalbumin (OVA) in 1 ml of saline with 100 mg of alu minum hydroxide. From day 15 the animals were challenged with aerosolized OVA (1% in saline) for 20 minutes per day for 7 consecutive days. (2) MT group: OVA-sensitized rats were injected intraperitoneally with 10 mg/kg MT 30 minutes before each OVA challenge. (3) CONTROL GROUP: OVA for inhalation and MT for intraperitoneal injection was replaced with normal saline (NS). Airway responsiveness to aerosolized acetylcholine of 24 rats was detected six hours after the last challenge. Then the rats were lavaged and total and differentiated leukocytes counts in bronchoalveolar lavage fluid (BALF) were performed after staining with Wright-Giemsa staining. At the same time, levels of nitric oxide (NO) in BALF, inducible nitric oxide synthesis (iNOS) and constitute nitric oxide synthesis (cNOS) in the lung tissues were assessed with the use of nitrate reductase and chemical colorimetry, respectively. The expression of NF-kappaB in the lung tissues was observed by means of immunohistochemical staining.

RESULTS(1) After OVA challenge, there was a significant decrease in airway responsiveness, lymphocytes and eosinophils in BALF in MT group compared with asthmatic group (P < 0.01 respectively); (2) There was a significant decrease in amounts of NO(2)(-)/NO(3)(-) in the BALF and levels of iNOS in the lung tissues in MT group comparing with asthmatic group (P < 0.01 respectively); and the levels of iNOS in the lung tissues was correlated positively with NO(2)(-)/NO(3)(-) in the BALF (P < 0.01), but there were no significant differences in activity of cNOS in any of the groups analyzed. (3) There was a significant increase in expression of NF-kappaB in lung tissues in asthmatic group compared with the other groups (P < 0.01), and so was in MT group compared with control group (P < 0.05).

CONCLUSIONSMT could partially inhibit the expression of NF-kappaB and down-regulate the activity of iNOS in lung tissue, decrease the production of NO in BALF. These data suggest that the inhibitory effect of MT probably play a role in decreasing airway hyperresponsiveness and airway inflammation of asthmatic rats model.

Animals ; Antioxidants ; pharmacology ; Asthma ; drug therapy ; metabolism ; Bronchoalveolar Lavage Fluid ; cytology ; immunology ; Disease Models, Animal ; Lung ; chemistry ; pathology ; Male ; Melatonin ; pharmacology ; NF-kappa B ; analysis ; Nitric Oxide ; analysis ; Rats ; Rats, Sprague-Dawley

Tumor angiogenesis provides adequate oxygen and nutrition for tumor development and supports tumor growth and metastasis. Stromal cell derived factor 1 (SDF-1) and its receptor C-X-C motif chemokine receptor 4 (CXCR4) in pancreatic cancer microenvironment are involved in tumor growth such as promoting tumor cell proliferation, migration, and angiogenesis. In this study, anti-CXCR4 nanobody (CXCR4 Nb) and anti-programmed cell death ligand 1 (PD-L1) &CXCR4 bispecific nanobody (PX4 BsNb) were expressed in Escherichia coli system and purified by nickel column affinity chromatography. We investigated the anti-angiogenesis activity and mechanism of CXCR4 Nb by in vivo and in vitro experiments. Ethical approval was obtained for collection of human peripheral blood mononuclear cell (hPBMC) samples from the Local Ethics Committee of Shanghai Jiao Tong University. All animal experiments were approved by the Animal Ethic Committee of Shanghai Jiao Tong University. The results showed that CXCR4 Nb at 0.1 μmol·L^-1 could effectively inhibit the proliferation and migration of human umbilical vein endothelial cells (HUVEC) promoted by pancreatic stellate cells in vitro. CXCR4 Nb and PX4 BsNb at 0.3 mg·kg^-1 obviously decreased tumor angiogenesis and inhibited the tumor growth in NOD/SCID mice, the inhibitory rates were 28.8% and 36.1%, respectively. CXCR4 Nb significantly inhibited tumor growth and angiogenesis with great safety, which provides support for application of CXCR4 Nb and anti-angiogenesis therapy of pancreatic cancer.

OBJECTIVETo measure the utility value of different skeletal malocclusion for patients receiving orthodontic treatment.

METHODSUtility value of different skeletal malocclusion for patients was measured by rating scale and time trade-off.

RESULTSThe youth group had higher utility values than adult group for skeletal malocclusion Class II (protruding facial type) with median mandibular angle. The utility value of skeletal malocclusion Class III (concave facial type) with low mandibular angle was the lowest, and the utility value of skeletal malocclusion Class II (protruding facial type) with median mandibular angle was the highest. There was no difference in the utility values by rating scale and by time trade-off.

CONCLUSIONFor some skeletal malocclusion, the youth had different utility values with the adult.

Adolescent ; Adult ; Cephalometry ; Face ; Female ; Humans ; Male ; Malocclusion ; Malocclusion, Angle Class III ; Mandible

OBJECTIVE: To analyze the clinical features of invasive fungal infection in patients with hematological malignancies and to compare the the therapeutic effect of fluconazole and intraconazole. METHODS: The clinical manifestations, mycological features, and the therapeutic results of 47 patients were retrospectively analyzed. Fluconazole was given to 17 paitents, intraconazole was given to 21 patients, and intraconazole to the other 9 patients after they had no effect with fluconazole. RESULTS: All patients had fever. The lung and the mouth cavity were the main locations of infection (53.2% and 21.3%, respectively). Fungi were found in 23 (48.9%) patients, in which the majority were Candida albicans and Aspergillus (56.5% and 26.1%, respectively). Intraconazole was more effective than fluconazole (63.3% vs. 34.6%, P<0.05) with no serious side effect. CONCLUSION The most common clinical features of IFI are fever, lung infection, and oral infection in patients with hematological malignancies. Candida albicans and Aspergillus infection are common. Intraconazole is safe and effective for invasive fungal infection.
Adolescent ; Adult ; Aged ; Antifungal Agents ; therapeutic use ; Aspergillosis ; complications ; diagnosis ; drug therapy ; Candidiasis ; complications ; diagnosis ; drug therapy ; Female ; Fluconazole ; therapeutic use ; Hematologic Neoplasms ; microbiology ; Humans ; Itraconazole ; therapeutic use ; Lung Diseases, Fungal ; complications ; diagnosis ; drug therapy ; Male ; Middle Aged

OBJECTIVETo compare the effect of the extracts from Decoction for resuscitation (DRE) and its component herbs on prostacyclin (PGI2), thromboxane A2 (TXA2) and nitric oxide (NO) release from rat vascular endothelial cells under hypoxia.

METHODAfter treatment with the extracts from DRE and its component herbs, the contents of 6-keto-prostaglandin F1alpha(6-keto-PGF1alpha), thromboxane B2 (TXB2) as well as nitrite (NO), which were degradation products of PGI2, TXA2 and NO respectively, in culture medium of rat vascular endothelial cells under hypoxia were measured with radioimmunoassay and Griess Reaction.

RESULTCompared with the control group, the results indicated that DRE, prepared licorice root extract (LE), dried ginger extract (GE), aconite root extract (AE), extracts of aconite root and prepared licorice root (ALE), extracts of aconite root and dried ginger (AGE) increased significantly the content of 6-keto-PGF1alpha and the ratio of 6-keto-PGF1alpha/TXB2, but had no effect on the content of TXB2 in culture medium of rat vascular endothelial cells under hypoxia. The content of 6-keto-PGF1alpha in the DRE group was higher than that in the groups of LE, GE, AE, ALE, AGE. The ratio of 6-keto-PGF1alpha/TXB2 in the DRE group was higher than that of the groups of GE, AE, ALE. Compared with the control group, DRE, LE, GE, AE, ALE, AGE increased significantly the content of NO2- in culture medium of rat vascular endothelial cells under hypoxia. Moreover, the content of NO2- in the DRE group was higher than that of the groups of GE, AE, ALE.

CONCLUSIONThe results suggested that DRE increased significantly the content of PGI2 and the ratio of PGI2/TXA2 as well as the content of NO. The effect of DRE on the parameters in culture medium of rat vascular endothelial cells under hypoxia was better than that of the extracts from its component herbs.

6-Ketoprostaglandin F1 alpha ; metabolism ; Aconitum ; chemistry ; Animals ; Aorta, Abdominal ; cytology ; Cell Hypoxia ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Endothelial Cells ; metabolism ; Ginger ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Nitric Oxide ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Thromboxane B2 ; metabolism

BACKGROUNDFamilial hypercholesterolemia (FH) is an autosomal disorder associated with elevated plasma low density lipoprotein (LDL) levels leading to premature coronary heart disease (CHD). As a result of long-term hyperlipemia, FH patients will present endarterium thickening and artherosclerosis. In the present study we scanned the related gene of a clinically diagnosed autosomal genetic hypercholesterolemia family for the possible mutations and established eukaryotic expression vector of mutation of proprotein convertase subtilisin/kexin type 9 (PCSK9) gene with gene recombination technique to investigate the contributions of the variation on low density lipoprotein receptor (LDL-R) metabolism and function alternation.

METHODSMutation detection was conducted for LDL-R, apolipoprotein B(100) (apoB(100)) and PCSK9 gene with nucleotide sequencing in a Chinese FH family. The full-length cDNA of wild type PCSK9 gene (WT-PCSK9) was obtained from Bel-7402. Site mutagenesis was used to establish the recombinant eukaryotic expression vector carrying pathogenic type of PCSK9 gene and the inserted fragment was sequenced. With the blank vector as control, liposome transfection method was used to transfect the Bel-7402 cells with recombinant plasmid. The expression of LDL-R mRNA was examined by RT-PCR. PCSK9 and the expression of LDL-R protein were determined by Western blotting.

RESULTSThe G-->T mutation at the 918 nucleotide of PCSK9 gene resulted in the substitution of the arginine by a serine at the codon 306 of exon 6. After sequencing, it was confirmed that the inserted fragment of established expression vector had correct size and sequence and the mutant was highly expressed in Bel-7402 cells. There was no significant variation in the levels of LDL-R mRNA. LDL-R mature protein was decreased by 57% after the cells were transfected by WT-PCSK9 plasmid. Mature LDL-R was significantly decreased by 12% after the cells were transfected by R306S mutant as evidenced by gray scale scanning, suggesting that the new mutant R306S can significantly decrease the expression of mature LDL-R protein.

CONCLUSIONSA novel missense mutation of PCSK9 gene, R306S, was found and the eukaryotic expression vectors of mutant and wild-type of PCSK9 gene were established. There was no significant variation in the levels of LDL-R mRNA. The R306S mutation could significantly lead to the decrease of LDL-R mature protein expression, which might be the pathogenic gene of the FH family.

Adolescent ; Adult ; Female ; Humans ; Hyperlipoproteinemia Type II ; genetics ; Lipids ; blood ; Male ; Mutation ; Pedigree ; Proprotein Convertase 9 ; Proprotein Convertases ; Serine Endopeptidases ; genetics