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 detect molecular characterization of the promoter and 5′UTR region of ATP7B gene in Chinese children with Wilson disease (WD) and explore the distribution of polymorphisms and mutations in different ethnicity.Methods One hundred and ten patients with WD and 90 healthy children were enrolled into the study and analyzed by polymerase chain reaction-single strand configuration polymorphism (PCR-SSCP) and DNA sequence analysis.Results 1.Five polymorphisms were identified as follows, -1294T→G,-105C→G,-116C→T ,-132delGCCGC and -75A→C(transcription start site as +1). The former three ones had never been reported before. The fourth one had not been reported either in China. 2.The polymorphism -132delGCCGC and -75A→C both exhibited with allelic frequency at above 70%, which was much higher than other races. The -132delGCCGC polymorphism shared almost complete linkage disequilibrium with the -75A→C polymorphism (in 98% patients) and their regularity was 96.9%.3. Almost all the polymorphisms distributed in flanking sequence of EXON 1 in Chinese. Race and geological distribution may be dominant factors of the variability of promoter and 5′UTR region of ATP7B gene.Conclusions Three novel polymorphisms and a linkage disequilibrium with the -132delGCCGC and -75A→C were identified in Chinese with WD. It also suggests that the mutation in the promoter of ATP7B is uncommon in Chinese patients.

Ataxia Telangiectasia Mutated Proteins ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Cell Cycle Proteins ; metabolism ; Checkpoint Kinase 2 ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Neoplasm Grading ; Protein-Serine-Threonine Kinases ; metabolism ; Tumor Burden ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism

This paper introduced a composite method of treating gastrointestinal tract cancer-thermo-chemotherapy.A complete set of intelligentialized real-time measure and control system was well designed. The experiment result showed that this system has a stable performance and good real-time.

Objective To investigate the relationship between matrix metalloproteinase(MMP)1 gene-519A/G polymorphism and the risk of coronary heart disease(CHD)in Northern Chinese Han population.Methods A total of 517 patients with CHD and 380 healthy adults diagnosed by coronary angiography were genotyped by polymerase chain reaction-restriction fragment length polymorphism and DNA sequence technology for the-519A/G polymorphism in MMP1 gene.Results (1)The frequency of AA genotype was significantly higher in patients with CHD than that in controls[67.70%(350/517) vs.40.26%(153/380),OR=1.64,P＜0.001,95%CI:1.44-1.86].People carrying A allele had increased risk for CHD(OR=1.49,P＜0.001,95% CI:1.33-1.69).(2)The frequency of AA genotype was higher in patients with acute coronary syndrome(ACS)than patients with stable angina pectoris[68.81%(278/404)vs.51.76%(44/85,),P＜0.01,95% CI:1.04-1.27].The A allele carriers were more likely to develop ACS(OR=1.11,95% CI:1.01-1.21,P＜0.05).Conclusion Our data shows MMP1 gene -519A/G polymorphism is associated with the risk of CHD,and A allele carriers are more susceptible for CHD in Northern Chinese Han population.

OBJECTIVETo classify NZB/W F1 lupus mice into three different constitutions, i. e. the cold, normal, and hot constitutions, and to prove the objective existence of the difference among them.

METHODSUsing the Four Diagnosis Work Station for Mice (founded by Prof. FANG Zhao-qin, the Research Faculty of Experimental Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine), the body weight, the armpit temperature, the heart rate, the 35-s activities, and the color of their tails and claws (r value) were detected. The total weight was calculated according to the formula: The total weight sum = the correction value of claws x 1 + the correction value of tail x 0.2 + the correction value of the armpit temperature x 0.7 + the correction value of heart rate x0. 05 + the number of the quadrant crossing x 0.025. The NZB/W F1 lupus mice were classified into the three different constitutions according to the higher total weight sum, the hotter the constitution, the lower total weight sum, the colder the constitution. The hydroxyproline content, connective tissue growth factor (CTGF), and transforming growth factor-beta1, (TGF-beta1) gene expression difference in the renal tissue were detected and the immunofluorescence staining observed.

RESULTSAmong the 158 NZB/W F1 lupus mice, 33 mice were classified into the hot constitution, 34 into the cold constitution, and 91 into the normal constitution. The renal hydroxyproline content in mice of normal and cold constitutions were higher than that of mice of the hot constitution (P<0.01). No statistical difference was shown between mice of the normal constitution and mice of the cold constitution. The CTGF gene expression level was significantly higher in mice of the cold constitution and mice of the hot constitution than in mice of the normal constitution (P<0.05). No significant difference was found between mice of the cold constitution and mice of the hot constitution. Lower level of TGF-beta1, expression existed in mice of the cold constitution than in mice of the normal constitution or mice of the hot constitution, showing insignificant difference. The immunofluorescence stain of the renal tissue among the three constitutions also showed some difference.

CONCLUSIONConstitution difference did exist among NZB/W F1 lupus mice.

Animals ; Body Constitution ; Disease Models, Animal ; Female ; Kidney ; metabolism ; pathology ; Lupus Erythematosus, Systemic ; metabolism ; pathology ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred NZB ; classification

To construct knockout vectors containing ampicillin resistant gene and partial sequence of hyaluronidase gene(Hyl)so that Hyl can be knock out by transforming the plasmid into Streptococcus zoopidemics mutans.First,partial sequence of Hyl(Hyl-1)was cloned into the vector of pMD19-T by using DNA of Streptococcus zoopidemics as template,and then a knockout vector pMD19T-SA was constructed,in which Hyl-1 gene was disrupted by inserting ampicillin resistant gene(Amp)from reverse PCR.As expected,the vector was proved to be consisted of Hyl-1-Amp-Hyl-1-pMD19-T.Thereafter,DNA fragment of Hyl-1-Amp-Hyl-1 was subcloned into pBR322 vector,the resulting construct was then checked by PCR and restriction analysis for the proper configuration of the knockout vector pBR322-SA.Both of the knockout vectors were used to transform Streptococcus zoopidemics and one recombinant was obtained in result.From results of PCR and Hyl activity assay,it was indicated that in the recombinant the Hyl gene was disrupted completely.

This study is to establish an HPLC-DAD-ELSD method for simultaneous determination of 5 flavones and saponins in Rhizoma Anemarrhenae including neo-mangiferin, mangiferin, timosaponin B II, timosaponin B III and timosaponin A III. Samples were analyzed on a Merck Purospher STAR column(4.6 mm x 250 mm, 5 μm). The mobile phase consisted of acetonitrile( A) and 0. 1% formic acid (B) with gradient elution at a flow rate of 1.0 mL · min(-1). The column temperature was set at 40 °C. The DAD detector wavelength was set at 254 nm. The ELSD conditions were as follows: the nebulizing gas flow rate was 2.0 L · min(-1) and temperature of drift tube was 105 °C. The volume was 10 μL. The five compounds were well separated with good linear correlations. The mean recoveries were between 102.0%-104.0%. This method was quick and reliable which provides a foundation for quality control of R. Anemarrhenae.
Anemarrhena ; chemistry ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; analysis ; Flavones ; analysis ; Rhizome ; chemistry ; Saponins ; analysis

Objective:To explore the relationship among psychological distress,adult attachment,and social support in primary caregivers of cancer patients.Methods:A total of 208 primary caregivers of cancer patients in one third grade hospital in Anhui Province were recruited.The 10-item Kessler Psychological Distress Scale(K10),Experiences in Close Relationships Inventory(ECR) and Social Support Questionnaire(SSQ) were used to explore psychological distress,adult attachment,and social support status.Results:The average K10 score was (21.5 ± 7.5).Multiple linear regression analysis indicated that caregiver gender,pressure on patient care,and attachment anxiety had positive prediction on psychological distress (β =2.30,3.02,0.13),while residence had negative prediction on psychological distress (β =-3.22).Conclusion:It suggests that the psychological distress is related to attachment anxiety and social support among the primary caregivers.

Objective To screen the mutation of certain gene of a 10-years-old boy with multiple xanthomas and very high level of cholesterol who could be diagnosed as homozygous familial hypercholesterolemia (FH),to explore the relationship between the genotype and phenotype,and to discuss the molecular pathologic mechanism.Methods The basic information of life styles were asked from the boy and his familial members.The blood was drown to examine the lipid and genes.The boy was examined with electrocardiogram examination,ultrasonography and coronary CT angiography (CTA) to evaluate the degree of atherosclerosis.Peripheral blood DNA of the boy and his parents were extracted by phenol-chloroform method and investigated for mutations of promoter and all 18 exons of low density lipoprotein receptor(LDLR) gene.Screening was carried out by using Touch-down polymerase chain reaction (PCR) and single strand conformation polymorphism(PCR-SSCP),combined with DNA sequence analysis.In addition,the apolipoprotein B100 gene(apoB100) for known mutations (R3500Q) which caused familial defective apoB100 was screened by PCR-DNA sequence analysis.Results 1.The level of cholesterol of his parents were higher than the normal.2.Several clinical manifestations of atherosclerosis were detected from that boy.Increased intima-media thickness and plaques were detected in the common carotid artery.Mitral valve regurgitation was found by echocardiography.Coronary stenosis was confirmed by CTA.3.No mutations R3500Q of apoB100 was observed.4.A homozygous mutation in exon13 of the LDLR gene (D601Y) were identified in the boy and his parents harbour D601Y heterozygous mutation due to a single base pair substitution of G for T in the codon for residue 1864.Conclusions The final diagnosis of the boy with multiple xanthomas was homozygous FH.His disease was caused by D601Y homozygous mutation in exon13 of the LDLR gene inherited from his heterozygous parents.