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

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 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.

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

Objective:To investigate the relationship between angiopoietin-like protein 4(ANGPTL4) and vascular endothelial growth factor(VEGF) in serum and lower limb arterial lesions in Type 2 diabetes mellitus.Methods:Using the method of retrospective cohort study, 164 inpatients with type 2 diabetes admitted to the Department of Endocrinology, Dahua hospital, Xuhui District, Shanghai from October 2017 to December 2018 were selected as the type 2 diabetes group, and 69 healthy people in the same period were collected as the normal control group.According to the ankle brachial index<0.9, diabetic patients were divided into two groups: the group without lower extremity artery disease and the group with lower extremity artery disease.The levels of ANGPTL4 and VEGF in serum were measured by enzyme-linked immunosorbent assay.Results:(1)Compared with the normal control group: the concentration of ANGPTL4 in serum of lower extremity arterial disease group and no lower extremity arterial disease group((44.78±15.15), (47.46±10.43) μg/L) were lower than that of normal control group((52.87±12.74) μg/L), the difference was statistically significant (all P<0.05). The serum VEGF concentration in lower extremity arterial disease group and no lower extremity arterial disease group((340.98±76.18), (314.83±75.30) ng/L) were higher than that in normal control group((282.58±81.4) ng/L), the difference was statistically significant(all P<0.05). (2)Compared with no lower extremity arterial disease group: the concentration of ANGPTL4 in serum of lower extremity arterial disease group was lower than that of no lower extremity arterial disease group, but the difference was not statistically significant( P>0.05). The serum VEGF concentration in lower extremity arterial disease group was higher than that in no lower extremity arterial disease group, the difference was statistically significant( P<0.05). (3)Correlation analysis: Diabetes mellitus and diabetic course>10 years( OR 4.594, 95% CI2.540-8.311, P<0.001), fasting blood glucose>8 mmol/L( OR 2.353, 95% CI1.023-5.416, P=0.044), glycosylated hemoglobin>8%( OR 6.442, 95% CI 2.874-14.441, P<0.001) and the increase of VEGF( OR 3.745, 95% CI1.566-8.953, P=0.003) are risk factors for lower limb arterial disease in type 2 diabetes, while the increase of ANGPTL( OR 0.154, 95% CI0.064-0.372, P<0.001) can reduce the occurrence of lower limb arterial disease in type 2 diabetes. Conclusion:ANGPTL4 is a protective factor and VEGF is a risk factor of type 2 diabetes mellitus.Both of them influence each other and participate in its occurrence and development.

Objective:To explore the relevant factors for remote diffusion-weighted imaging lesions (R-DWILs) in patients with spontaneous intracerebral hemorrhage.Methods:Two hundred and three patients with spontaneous intracerebral hemorrhage, admitted to our hospital from January 2018 to December 2019, were enrolled. According to the presence of R-DWILs, these patients were divided into positive group ( n=39) and negative group ( n=164). The basic information of R-DWILs in the positive group was analyzed. The basic demographic and clinical data and laboratory examination results were collected and compared between the two groups. Subsequently, the items with P<0.1 were included in the multivariate Logistic regression analysis to identify the independent influencing factors for R-DWILs. Results:A total of 55 R-DWILs were detected in 39 patients (19.2%) with spontaneous intracerebral hemorrhage, including 45 in the cortical and subcortical area (81.8%), 8 in the basal ganglia area (14.5%), one in the brainstem area (1.8%), and one in the cerebellum area (1.8%). The diameter of these lesions ranged from 2-20 mm, which were round or oval, flaky, irregular and so on. Forty-two R-DWILs (76.4%) were in the contralateral hemisphere, and the number of R-DWILs was 1-3 in these patients. Systolic blood pressure, diastolic blood pressure, white blood cell count (WBC), neutrophil count, neutrophil to lymphocyte ratio (NLR), fasting blood glucose level in the positive group were significantly higher than those in the negative group ( P<0.05). Logistic regression analysis showed that fasting blood glucose level ( OR=1.632, 95%CI: 1.213-2.072, P=0.015), neutrophil count ( OR=1.037 , 95%CI: 1.019-1.581, P=0.042), and NLR ( OR=2.151 , 95%CI: 1.397-2.733, P=0.003) were independent influencing factors for R-DWILs. Conclusions:R-DWILs are common in patients with spontaneous intracerebral hemorrhage, the lesions mainly occur in the cortical and subcortical area, and it is more common in the contralateral hemisphere. Patients with high fasting blood glucose, neutrophil count, and NLR are trended to have R-DWILs.

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