Objective To analyze the epidemiological characteristics of Mycobacterium tuberculosis by enterbaeterial repetitive intergenic consensus-polymerase chain reaction(ERIC-PCR)DNA fingerprint. Methods Mycobacterium tuberculosis positive sputum samples between September 2003 to May 2006 were collected and cultured.Chromosomal DNA were extracted and ERIC-PCR DNA fingerprinting was analyzed by software,such as RAPD PHYLIP and Treeview.Results A total of 42 different fingerprints were detected.Phylogenetic analysis showed that they could be classified into three clusters,the clustering rate was 72.6%.The characteristics of ERIC-PCR fingerprint patterns were related to age,drug resistance,and type of resistance.Conclusions ERIC-PCR DNA fingerprinting technique used in this study is good for epidemiological studies with its strong discrimination,simplicity and rapidness.A high level of recent transmission is found in our city.

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.

Health Personnel ; psychology ; Humans ; Logistic Models ; Multivariate Analysis ; Professional Competence ; statistics & numerical data ; Stress, Psychological ; epidemiology

With the rapid development of medical genetics,online Medelian inheritance in man (OMIM) manifests a more and more important role in medical genetics teaching.Using the educational form combining ‘ classroom teaching,review writing and seminar’,‘ Query and use of OMIM ’was introduced into the education of medical genetics.Reality practice revealed that this educational practice maintained advanced and timely status of knowledge and deeply activated self-studying and independent thinking ability of students.

BACKGROUND:Rapid loss of liver-specific functions of the cultured hepatocytes limits the development of hepatocyte-based therapies. OBJECTIVE:To establish a three-dimensional culture system based on col agen hydrogel that enables to enhance liver-specific functions for a long period during culture of hepatocytes. METHODS:Hepatocytes were isolated from Sprague-Dawley rats and then encapsulated into liquid type Ⅰcol agen solution that was premixed with hepatocyte growth factor and Dulbecco's modified Eagle’s medium to create hepatocyte/col agen hydrogel compounds. The compounds were inoculated into a 96-wel plate. After gelation, culture medium was added. Light microscope, hematoxylin-eosin staining and transmission electron microscopy were used to examine the morphological characteristics and ultrastructure of the cultured hepatocytes. The cellsupernatant was col ected and tested for albumin secretion and urea synthesis. Periodic acid-schiff staining, immunofluorescence staining and quantitative real-time PCR were also used to further clarify liver-specific phenotype or function of the hepatocytes.RESULTS AND CONCLUSION:(1) Light microscope revealed that hepatocytes were round shape and distributed uniformly in col agen hydrogel. The three-dimensional hepatocyte culture system exhibited similarities to liver-like structure and tight junction were formed between hepatocytes after 14 days of culture. (2) Within the three-dimensional culture system, hepatocytes remained positive for periodic acid-schiff staining, albumin and hepatocyte nuclear factor-4αpositive after 14-day culture, which provided the convincing evidence of highly differentiated primary hepatocytes with functions of glycogen and albumin synthesis. (3) The albumin and urea productions in the three-dimensional culture system had a significantly higher level than in the two-dimensional culture, and could remain at a high level at least for 15 days. (4) The expression levels of hepatocyte-specific genes including Albumin, HNF-4α, Claudin-3, CYP1A1, CYP3A1 and G6P were significantly improved in the three-dimensional culture as compared with the two-dimensional culture. The col agen hydrogel based three-dimensional culture system provides a valuable model to enhance the hepatocyte functional maintenance and lay the foundation for the development of hepatocyte-based therapy for liver disease.

Female ; Humans ; Infant, Newborn ; Maple Syrup Urine Disease ; diagnosis ; therapy

OBJECTIVETo investigate the protective effect of epimedium flavonoids Injection (EFI) on experimental acute myocardial infarction (AMI) rats.

METHODSRats were randomly divided into 6 groups, the acute myocardial infarction model was established by ligating left anterior descending branch of coronary artery (LAD). After operation, the rats in the sham-operation and model group were intravenous injected with 5% glucose injection, those in the positive medicine group were intravenous injected with nitroglycerin 0.3mg/kg, while rats in the low-, middle- and large-dose EFI group were intravenous injected with TFE in a dose of 10, 20, 40 mg/kg respectively. ECG was monitored before and after coronary artery ligation, and after treatment at different time points. At the same time, the millivolt of ST and ST-T segment were measured. The changes of serum creatine phosphokinase (CPK), lactate dehydrogenase (LDH), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were determined, and the myocardial infarcted area was detected by MTT respectively 3 h after LAD. Results After intravenous injection of EFI in a dose of 10, 20, 40 mg/kg, the myocardial infarcted area of AMI rats could be decreased in different degree, the activity of serum CPK, LDH and the content of MDA decreased (P < 0.05 or P < 0.01), while the activity of serum SOD increased significantly (P < 0.05 or P < 0.01). It could began to lower the elevated ST-T segment 5 min after medication and the action could last for 3 h (P < 0.05 or P < 0.01).

CONCLUSIONEFI has a protective effect against acute myocardial ischemia caused by LAD, and the effect is quickly initiated.

Animals ; Creatine Kinase ; blood ; Epimedium ; chemistry ; Female ; Flavonoids ; isolation & purification ; therapeutic use ; L-Lactate Dehydrogenase ; blood ; Male ; Myocardial Infarction ; drug therapy ; Phytotherapy ; Random Allocation ; Rats ; Rats, Wistar ; Superoxide Dismutase ; blood

OBJECTIVETo explore the clinical anesthesia value of transcutaneous acupoint electrical stimulation (TAES) combined with general intravenous anesthesia in endoscopic bilateral thyroidectomy patients.

METHODSTotally 60 patients who underwent endoscopic bilateral thyroidectomy were equally randomly assigned to 2 groups, the treatment group and the control group, 30 in each group. Patients in the treatment group received TAES combined general intravenous anesthesia, while those in the control group received total intravenous anesthesia. Anesthesia was maintained by target controlled infusion of propofolum combined constant speed infusion of remifentanil in the two groups. TAES was maintained from 30 min before anesthesia induction to the end of endoscopic thyroidectomy at bilateral Hegu (L14) and Neiguan (PC6). The mean artery pressure (MAP) and heart rate (HR) were recorded at different time points of anesthesia, i.e., immediately after entry into the operating room (TO), immediately after intubation (T1), 5 min after intubation (T2), 5 min before incision (T3), 5 min after incision (T4), 30 min after inflation (T5), at the end of surgery (T6), 5 min before extubation (T7), immediately after extubation 0 (T8), and 5 min after extubation (T9). The concentration of IL-6 and TNF-alpha were measured at TO, T3, T5, and T6. The target concentration of propofol was also recorded at T3, T4, and T5.

RESULTSThere was no statistical difference in the operation time between the two groups (P >0.05). Compared with TO in the same group, HR at T3-T4 decreased and increased at T8-T9, and MAP increased at T7-T9 in the treatment group; HR decreased at T3 and increased at T7-T9, MAP increased at T1, T5, T7-T9, and MAP decreased at T2-T3 in the control group. IL-6 increased at T5-T6, while TNF-alpha decreased at T6 in the two groups (P <0.01,P <0.05). Compared with the control group at the same time point, HR decreased at T6-T9, MAP decreased at T1, T4, T5, T7-T9, MAP increased at T3, and IL-6 decreased at T5-T6 in the treatment group (P <0. 05). The concentration and the total amount of propofol were significantly lower in the treatment group than in the control group (P <0.01,P <0.05).

CONCLUSIONSTAES could maintain the hemodynamics more stably and inhibit the stress response in endoscopic thyroidectomy. It also reduce the dosage of anesthetics and improve the safety of anesthesia.

Acupuncture Points ; Anesthesia, General ; Anesthesia, Intravenous ; Electric Stimulation ; methods ; Endoscopy ; methods ; Heart Rate ; Hemodynamics ; Humans ; Interleukin-6 ; Piperidines ; Propofol ; Thyroidectomy ; methods ; Transcutaneous Electric Nerve Stimulation ; Tumor Necrosis Factor-alpha

Objective To evaluate the performance of the ABX Micro C-reactive protein(CRP)in determination of CRP.Methods The analytic characteristics including precision,carry-over,linearity, stability,interference and comparability were examined.Results The coefficient of variation(CV)was less than 5.1%,10% and d.3% for within-run,between-run and between-day,respectively.Carryover was less than 1.2%.Whole blood samples held at either room temperature or 4℃ were stable for 48 hours with relative deviation less than 6.0% relatively.Linear range was 1.0-70.0 mg/L using undiluted samples.The comparison between the ABX Micro CRP and Behring Nephelometer Ⅱ was well correlated Both serum:Y=0.996 7X-0.398 5,r~2=0.965 9;serum for BN Ⅱ,whole-blood samples for the ABX Micro CRP:Y=0.908 8X-0.138 2,r~2=0.959 4;both serum and whole-blood samples for the ABX Micro CRP: Y=1.001 7X-0.898 2,r~2=0.952 7.No obvious interference was observed by hyperhemoglobinemia and hyperlipidemia.Conclusion The determination of CRP test with ABX Micro is accurate and reliable.

The aim of this study was to investigate the effect of Celastrol on induction of HL-60 cell apoptosis and its possible mechanism. The proliferative activity of HL-60 cells treated with 0.25 - 8.0 μmol/L of Celastrol for 24 - 72 hours was assayed by MTT method, the effects of Celastrol on apoptosis and cell cycle of HL-60 were detected by TUNEL staining and flow cytometry with Annexin V-FITC/PI double labeling, the expression of pAkt and cyclin D1 at protein and gene level in HL-60 cells treated with Celastrol were measured by Western blot and RT-PCR. The results showed that the Celastrol could obviously inhibit the proliferation of HL-60 cells in concentration-and time-dependent manners, the IC₅₀ value of Celastrol for 24 hours was 6.21 ± 0.242 μmol/L. The Celastrol concentration-dependently induced the apoptosis of HL-60 cells, accompanying with morphological changes of apoptotic cells, which may be related with arrest of cells in G₀/G₁ phase. The Celastrol suppressed the expression of pAkt and Cyclin D1 in HL-60 cells to a varying degree which showed obvious concentration-and time-dependent manners. It is concluded that the Celastrol inhibits the proliferation and induced the apoptosis of HL-60 cells. Its mechanism may be related with down-regulation of p-Act and cyclin D1 expressions.
Apoptosis ; drug effects ; Cyclin D1 ; metabolism ; Down-Regulation ; Gene Expression Regulation, Leukemic ; HL-60 Cells ; Humans ; Proto-Oncogene Proteins c-akt ; metabolism ; Triterpenes ; pharmacology