China ; Humans ; Hypoxia-Ischemia, Brain ; diagnosis ; therapy ; Infant, Newborn

Objective To study whether RHD gene exists multiple mRNA produced by alternative splicing. Method RhD mRNA was analyzed through reverse transcriptase PCR (RT-PCR) and cDNA sequencing in 4 individuals with different Rh phenotypes (CcDDEe、CCDDee、CcDDee and ccDdee). Results The individuals with different Rh phenotypes had identical and intricate RhD mRNA isolations, which included normal mRNA, and 4 other transcripts with deletions of exon 7, exon 9, exon 7 and 9, and exon 7 to 9. Those transcripts had the same sequences as normal RhD mRNA except exon(s) deletion, which showed RHD gene existed alternative splicing, and it happened in exons or introns 7-9 regions. Conclusion There are intricate and different cassette-exons RhD transcripts produced through alternatively spliced exons 7 to 9. And those RhD mRNA isolations might code proteins with different C-terminus.

Objective To establish a RHD genotyping method specific for the Chinese. Methods Six pairs of primers specific for most alleles found in the Chinese according to the records in NCBI GenBank, were designed, and a multi-tube sequence-specific primer PCR (PCR-SSP) method was established with a pair of internal control primer in each reaction. The method was evaluated with samples serologically determined and full length RHD sequenced from 89 Rh-negative, 28 D el, and 13 Rh-positive, weak D and partial D phenotype of Chinese Hans. Furthermore, 318 random samples from blood donors were genotyped and the results were compared with serological results of those samples. Results The PCR-SSP results were in concordance with serological results (100%) in all samples, and all RHD positive, D antigen negative alleles (or nonfunctional alleles) observed in the Chinese up to now could be detected or implicated, including D el phenotype especially D el allele existing in Rh-positive individuals (RHD/RHD1227A). This genotype was detected with a rate of 8/318, and allele frequency should be 0.012579 Conclusion Our method is rapid and easy, with high accuracy in the testing of the Chinese.

The whole length of exon 11 of BRCA1 was sequenced (total 3427 bp) in 59 patients and 10 healthy female blood donors. To allow a rapid determination of the different BRCA1 alleles, a sequence-specific primer PCR method (PCR-SSP) was established and was applied to 57 additional female donors. Finally, the full-length coding region of BRCA1 was analyzed through reversed-transcriptase PCR (RT-PCR) and cDNA sequencing (total 5554 bp) in one donor with wild-type allele and 2 patients with one or two mutated alleles. By genomic DNA sequencing, 5 homozygous polymorphisms were observed in 18 patients: 2201C>T, 2430T>C, 2731C>T, 3232A>G and 3667A>G All of them were previously observed in Caucasians, Malay and Chinese, but for the first time the mutations were found in one allele (GenBank AY304547). Twenty-six patients and 4 donors were heterozygous at these 5 nucleotide positions. The remaining 15 patients and 6 donors showed a sequence identical with the standard BRCA1 gene. Combined the PCR-SSP results and in a summary, 6 of 67 (9.0 %) healthy individuals were homozygous for the mutated allele, whereas 18 of 59 (30.5 %) breast cancer patients were homozygous. A Chi-square test showed a significant correlation between homozygous mutated BRCA1 allele and breast cancer. The cDNA sequencing showed that 2 additional mutations, 4427T>C in exon 13 and 4956A>G in exon 16, were found. A new BRCA1 allele, which is BRCAI-2201T/2430C/2731T/3232G/3667G/4427C/4956G (GenBank AY751490), was found in Chinese. And the homozygote of this mutated allele may implicate a disease-association in Chinese.

Background As a new tonometer,it is necessary to assess the clinical value of Icare rebound tonometer.Objective This study was to compare the intraocular pressure(IOP)values measured by Icare with that measured by GAT,and discuss the clinical value of leare rebound tonometer. Methods IOP measurement was performed on 152 eyes of 78 subjects with suspicious glaucoma,glaucoma,refractive error and normal examinnee by Icare and GAT respectively.The Icare IOP was measured firstly and then the GAT IOP was carried out with the 3-or 5-minute interval.The IOP values were compared between Ieare and GAT.This study was approved by Ethic Committee of Wuhan General Hospital of Chinese PLA.Written informed consent was obtained from each subject prior to this study. Results The mean IOP values of Icare and GAT were(19.16±5.03)mmHg and(18.41±4.52)mmHg respectively.The differences between Icare IOP and CAT IOP were less than or equal to 1 mmHg in 96 of 105 eyes(63.2%).The positive correlation was found between the Icare IOP and GAT IOP(r=0.940,P<0.01).The Ieare IOP was lower than that of GAT when IOPIcare<16 mmHg,however,the IOP of Icare were higher when IOPIcare≥6 mmHg;the IOP of Icare were higher than that of GAT in the total CCT range.The correlation coefficients of IOP of Icare or CAT with CCT were 0.341(P<0.01)and 0.333(P<0.01),respectively. Conclusion Compared with GAT,Icare is more feasible in clinic because it is practicable and reliable.

OBJECTIVETo dynamically observe the effect of Shenshuai Mixture (SM) on repair of kidney in acute renal failure (ARF) rats.

METHODSMale SD rats were divided into 4 groups randomly, the normal group, the SM group, the verapamil group and the model control group. Except those in the normal group were treated with normal saline without modeling, all remaining rats, after being made into ARF model by intra-muscular injection of glycerin, were treated with SM, verapamil and normal saline respectively via gastrogavage. Renal function, renal pathology, mRNA expression of epidermal growth factor (EGF) and protein expression of proliferating cell nuclear antigen (PCNA) were detected once every day from the 1st day to the 5th day. Results (1) BUN and Scr levels increased markedly 24 hrs after modeling, but the Scr level in the two treated groups was significantly lower than that in the model group (P < 0.05). Compared with that in the model group and the verapamil group, renal function was better in the SM group on the 3rd day (P < 0.01), and approach to normal level on the 5th day. (2) Renal pathological changes alleviated in every phase of ARF in the SM group than that in the model group, especially part of tubule regeneration could be seen on the 3rd day (metaphase), and renal structure was rehabilitated on the 5th day (convalescence), prior to those in the model group. (3) At the 3rd day expression of EGF mRNA and PCNA in tubule epithelial cell (TEC) increased remarkably in the SM group, higher than those in the model and verapamil group (P < 0.05).

CONCLUSIONSM could promote renal tissue regeneration and rehabilitation, and shorten the course of ARF through up-regulating mRNA expression of EGF in TEC.

Acute Kidney Injury ; drug therapy ; physiopathology ; Animals ; Drugs, Chinese Herbal ; therapeutic use ; Epidermal Growth Factor ; biosynthesis ; genetics ; Kidney ; physiopathology ; Male ; Phytotherapy ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effect of the experiential compound prescription Shenshuai compound medicine (SSCM) on acute renal failure (ARF) rats, and expose its mechanism.

METHODMale SD rats were allocated into four groups at random. Except normal group, the others were injected glycerin into the musculi to induce a model of ARF. At the same time, rats in normal and model groups were given a dose (10 mL x kg(-1)) of normal saline; rats in the other two groups were given verapamil (Vp) 40 mg x kg(-1) x d(-1) and SSCM 22.5 mg x kg(-1) x d(-1) respectively by gastric gavages. In this way they were killed at 24 h after injecting glycerin.

RESULTIn contrast with model group, SSCM and Vp could ameliorate the renal function of acute tubular necrosis (ATN) rats markedly, in a way protect the renal inherent cell ultrastructure such as tubular epithelial cell etc at ATN early-stage; especially SSCM could enhance NO contents in serum, and was reduce ET levels inplasma, evidently cut down TNF-alpha contents in serum, and was partly superior to Vp.

CONCLUSIONIt is indicated that SSCM can adjust thebalance of contract and stretch blood vessal active substance and clear away initiate inflammatory medium, consequently alleviate renal pathological changes, prevent and treat ARF.

Acute Kidney Injury ; blood ; chemically induced ; pathology ; Animals ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Endothelins ; blood ; Epithelial Cells ; ultrastructure ; Glycerol ; Kidney ; pathology ; physiopathology ; Kidney Tubules ; pathology ; Male ; Nitric Oxide ; blood ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

The study was purposed to analyze DNA and allele structure of the partial D phenotypes D(Va) and D(VI) of the Rhesus blood group in Chinese. Through polymerase chain reaction (PCR) and direct genomic DNA sequencing, the RHD gene was detected in three weak D individuals identified serologically. The results showed that among the three weak D individuals, one was identified as partial D phenotype D(Va) (Hus) type and genotyped DccEe; another two were testified as D(VI) III type and genotyped DCcee. Moreover, the breakpoints of the replaced region by RHCE in D(Va) (Hus) were 5' end of the exon 5 and 3' end of the intron 5, and there were 7 novel polymorphisms in intron 5: 23-25(GCA)2, 98G>A, 168-169insG, 205-206insT, 494-495insA, 1256-1257insC, 1347G>T. In conclusion the whole exon 5 and intron 5 are replaced by RHCE in D(Va) (Hus) detected in Chinese.
China ; Exons ; genetics ; Genotype ; Humans ; Introns ; genetics ; Phenotype ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System ; genetics ; Sequence Analysis, DNA

Adult ; Bone Neoplasms ; diagnosis ; surgery ; Humans ; Ilium ; Magnetic Resonance Imaging ; Male ; Osteoma, Osteoid ; diagnosis ; surgery ; Tomography, X-Ray Computed

Digital potentiometers have been used in medical instruments. This paper describes the structure and principle of a digital potentiometer, especially its interfacing with a single chip processor and its programming.
Computers ; Equipment Design ; Equipment and Supplies