This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.
Acetylcholinesterase ; genetics ; metabolism ; Cholinesterase Inhibitors ; analysis ; pharmacology ; HEK293 Cells ; Humans ; Indans ; analysis ; pharmacology ; Inhibitory Concentration 50 ; Kinetics ; Piperidines ; analysis ; pharmacology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection

Objective To improve the knowledge of primary gout in children. Methods Clinical data of a 12-year-old girl with primary gout was collected. Analysis of UMOD gene, REN gene and HNF-1βgene was performed using PCR and di-rect sequencing. Results The girl was admitted for 1-month history of left hallux pain accompanied with elevations of serum uric acid concentration and serum creatinine concentration. Several examinations showed serum uric acid/creatinine ratio was greater than 2.5. The fractional excretion of uric acid was 3.4%-6.6%. The X-ray showed that the proximal phalanxes of halluces were erosion. The diagnosis of renal biopsy was ischemic renal injury and chronic tubulointerstitial nephropathy. Blood uric acid concentrations of parents were normal, and the family history of gout was negative. Two single nucleotide polymorphisms (c.264C>T heterozygous and c.866-71 G>A heterozygous) in UMOD gene, 1 single nucleotide polymorphism (c.373+44C>G heterozygous) in REN gene, and 2 single nucleotide polymorphisms (c.100-50-49ins TCTG heterozygous and c.781-22T>C homozygous) in HNF-1βgene were detected. No pathological mutation was detected in these 3 genes. Conclusions This child is highly suspected to have primary gout caused by familial juvenile hyperuricemic nephropathy.

A new benzaldehyde, 3-hydroxy-4-(4-(2-hydroxyethyl) phenoxy) henzaldehyde(1), together with six known compounds, including isovanillic acid(2), pyrocatechol(3), glutinosalactone A(4), chrysoeriol(5), apigenin(6) and luteolin(7) were isolated from aerial part of Rehmannia glutinosa. The compounds were isolated by macroporous resin, silica gel, Sephadex LH-20 and HPLC chromatographies. The chemical structures of 1-7 were elucidated on the basis of spectral analysis (MS, 1D NMR and 2D NMR).
Benzaldehydes ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Rehmannia ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Objective To evaluate the effect of computer-aided design of composite materials with epoxide acrylate maleic (E) and hydroxyapatite (H) in cranioplasty. Methods A total of 45 patients with cranium defects were treated with cranioplasty by using skull bone flaps made of composite materials including epoxide acrylate maleic (E) and hydroxyapatite (H) ,which was designed with computer aid according to individual requirements. The patients were followed up for 6-36 months. Results After cranioplasty with composite EH, there occurred subcutaneous fluid in one patient and mild bone collapse in one. The composite EH showed good histocompatibility, with no infection or rejection. Conclusion During cranioplasty, use of computer-aided design of composite EH takes advantages of good accuracy, short operation time, good biocompatibility and good clinical efficiency.

The efficacy and safety of uric-acid-lowering therapy (UALT) on slowing the progression of chronic kidney disease (CKD) accompanied by hyperuricemia were assessed. We searched Cochrane Library, PubMed, EMbase, CNKI, Wanfang and Vip databases up to November 15, 2012 for randomized controlled trials (RCTs) which compared the effect of UALT to control therapy in hyperuricemic patients secondary to CKD, and then performed quality evaluation and meta-analysis on the included studies. Seven RCTs involving 451 cases were included. UALT delayed the increase of serum creatinine (MD=-62.55 μmol/L, 95% CI: -98.10 to -26.99) and blood urea nitrogen (MD= -6.15 mmol/L, 95% CI: -8.17 to -4.13) as well as the decrease of glomerular filtration rate [MD=5.65 mL/(min·1.73 m2), 95% CI: 1.88 to 9.41], decreased systolic blood pressure (SBP) (MD= -6.08 mmHg, 95% CI: -11.67 to -0.49), and reduced the risk of the renal disease progression (RR=0.30, 95% CI: 0.19 to 0.46). However, there was no statistically significant difference in 24-h urinary protein quantity and diastolic blood pressure (P>0.05). We identified that UALT could delay the progression of CKD with secondary hyperuricemia. And this also indirectly proved that hyperuricemia was a risk factor for the CKD progression.

OBJECTIVE: To investigate the polymorphism of 11 canine STR loci. METHODS: A fluorescent multiplex system with 11 STR loci (PEZ1, PEZ2, PEZ3, PEZ5, PEZ6, PEZ8, PEZ12, FH2010, FH2054, FH2132 and FH2611) was constructed independently and performed to amplify 105 samples from dogs. The character of these loci was analyzed with the PCR data. RESULTS: The distributions of genotypes and allele frequencies of 11 STR loci were obtained. The total power of discrimination for the 11 loci in canine population was 0.9999999 and the cumulative probability of exclusion was 0.9330621. The observed heterozygosity and polymorphism information content (PIC) were 0.502 and 0.640, respectively. CONCLUSION Each of the eleven canine STR loci has a high genetic polymorphism and can be applied for the parentage testing and individual identification. The fluorescent multiplex system is a reliable method in forensic application.
Alleles ; Animals ; Dogs/genetics* ; Forensic Genetics ; Gene Frequency ; Genotype ; Microsatellite Repeats/genetics* ; Polymorphism, Genetic

OBJECTIVETo investigate the role of mutation and mRNA expression of tumor suppressor gene phosphatase and tensin homolog deleted in chromosome 10 (PTEN) in tumorigenesis and progression of oral squamous cell carcinoma (OSCC).

METHODSThe mutation of exon 3, 5, 6 and exon 8 of PTEN gene in 42 oral squamous cell carcinoma tissue and paired normal tissue were detected by polymerase chain reaction (PCR) and DNA sequencing methods. The levels of PTEN mRNA expression in these tissues were assayed using semi-quantitative RT-PCR. Data was analyzed with SPSS 14.0 software package.

RESULTSMutated exon 5 of PTEN gene was found in 2 cases of advanced OSCC. The expression of PTEN mRNA was detected in all OSCC and paired normal tissue. The level of PTEN mRNA in OSCC tissue (0.36 +/- 0.12) was significantly lower than that of paired normal tissue (0.64 +/- 0.09, P < 0.01).

CONCLUSIONSThe decreased expression of PTEN mRNA contributes to tumorigenesis of OSCC.

Carcinoma, Squamous Cell ; genetics ; Chromosomes, Human, Pair 10 ; Exons ; Gene Expression ; Genes, Tumor Suppressor ; Humans ; Microfilament Proteins ; Mouth Neoplasms ; genetics ; Mutation ; PTEN Phosphohydrolase ; genetics ; Phosphoric Monoester Hydrolases ; genetics ; Polymerase Chain Reaction ; RNA, Messenger ; metabolism ; Tensins

OBJECTIVETo explore pathogenesis of post-transplantation diabetes mellitus (PTDM) in renal transplantation recipients.

METHODSA total of 40 renal transplantation recipients were divided into three groups based on oral glucose tolerance test results: normal glucose tolerance (NGT) group (n = 10), impaired fasting glycaemia + impaired glucose tolerance (IFG + IGT) group (n = 16), and PTDM group (n = 14). Insulin resistance (IR) and beta cell function were assessed by homeostasis model.

RESULTSThe differences of the immunosuppressive agents used in these groups were not statistically significant (P > 0.05). Compared with NGT group, insulin area under curve and homeostasis model assessment-insulin resistance index were significantly higher in IGT + IFG group and PTDM group (P < 0.05). Compared with NGT group and IGT + IPG group, insulin secretion index at 30 min and homeostasis model assessment-insulin secretion index were significantly lower in PTDM group (P < 0.05).

CONCLUSIONInsulin resistance and beta-cell dysfunction may play a key role in the pathogenesis of PTDM.

Adult ; Diabetes Mellitus ; etiology ; physiopathology ; Female ; Humans ; Insulin Resistance ; Insulin-Secreting Cells ; physiology ; Kidney Transplantation ; Male ; Middle Aged ; Postoperative Complications ; etiology ; physiopathology ; Retrospective Studies ; Young Adult

OBJECTIVETo identify the risk factors of cardiovascular diseases and cerebrovascular diseases (CVD) events in kidney allograft recipients.

METHODSWe followed up 361 renal transplant recipients who had undergone renal transplantation in our center from January 2000 to December 2003 and evaluated the cumulative incidences and mortalities of CVD complications at baseline and post-transplantation 1, 3, 6, 12, 24, 36, 48, and 60 months. Kaplan-Meier plot was used to assess the incidence and Cox's proportional hazards model to determine the risk factors for cardiovascular complications.

RESULTSThe cumulative incidences of CVD were 3.1%, 5.4%, 9.9%, 13.0%, 18.0%, 21.1%, and 24.1%, 1, 6, 12, 36, 48, and 60 months after transplantation, respectively. History of diabetes mellitus (RR 2.19, 95% CI 1.32-3.97, P = 0.009) and CVD (RR 6.34, 95% CI 3.76-14.60, P = 0.002) as well as the post-transplantation hypertension (RR 1.18, 95% CI 1.02-1.34, P = 0.04), diabetes mellitus (RR 2.82, 95% CI 1.33-7.26, P = 0.002), hyperlipidemia (RR 2.04, 95% CI 1.26-5.17, P = 0.008) and abnormal creatinine (> 200 micromol/L, RR 1.81, 95% CI 1.08-3.21, P = 0.03), and proteinuria (> 0.3 g/d , RR 1.56, 95% CI 1.12-3.54, P = 0.05) were independently correlated with the development of cardiovascular events.

CONCLUSIONHistory of diabetes mellitus and CVD, post-transplant hypertension, diabetes mellitus, hyperlipidemia, abnormal creatinine and proteinuria are the independent risk factors of the development of CVD events.

Adolescent ; Adult ; Cardiovascular Diseases ; etiology ; Female ; Follow-Up Studies ; Humans ; Kidney Transplantation ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Risk Factors ; Young Adult

To evaluate the efficacy of an ultrasound-based quantitative method to diagnose liver fibrosis using a rat model. Ultrasonography was performed on the livers of 90 Sprague-Dawley rats with or without thioacetamide-induced fibrosis. The liver capsule thickness and 13 texture parameters of gray level co-occurrence matrix were extracted from the standard sonograms. After sacrifice, severity of liver fibrosis (S0-S4 classification) was diagnosed by histopathology. Analysis of variance and correlation statistical tests were used to analyze the differences between groups and determine the relationships between each of the 14 quantitative ultrasound index points and the histological results, respectively. Discriminant analysis models were developed for quantitative diagnosis of liver fibrosis, and the leave-one-case-out method was used to verify the efficiency of models. All 14 indices were significantly correlated with the histological stages of fibrosis (P less than 0.05). The accuracy of the discriminant model for S0, S1, S2, S3 and S4 was 83.3%, 84.2%, 70.0%, 50.0% and 88.2%, respectively. In addition, 73.3% of cross-validated rats were accurately classified. Grouping S0 as no fibrosis, S1 as mild fibrosis, S2 with S3 as moderate to severe fibrosis and S4 as early cirrhosis increased the accuracy of the discriminant model for these four groups (respectively, 91.7%, 84.2%, 69.0% and 88.2%) and allowed for 78.9% of cross-validated rats to be correctly identified. Ultrasonography combined with texture analysis was a novel and accurate method to diagnose liver fibrosis in a rat model; further studies may provide insights into its applicability for quantitating liver fibrosis in other animal models or in clinic.
Animals ; Liver ; diagnostic imaging ; pathology ; Liver Cirrhosis, Experimental ; diagnostic imaging ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Ultrasonography