In this paper are described the construction of biological valves poppet and the establishment of mathematical model according to cardiac valves hydrokinetics and the theory of membrane. Finite elements of biological valves leaflets connected with biological valves poppet are analyzed. The method of constructing geometric model is reasonable. CAD of biological valves poppet is realized by structured programming and parameterization.
Bioprosthesis ; Computer Simulation ; Computer-Aided Design ; Finite Element Analysis ; Heart Valve Prosthesis ; Humans ; Models, Theoretical ; Prosthesis Design ; methods

OBJECTIVETo construct an eukaryotic expression vector of human telomerase reverse transcriptase (hTERT) gene specific shRNA, and investigate the effect of pshRNA-hTERT combined with gamma-irradiation on cell survival and telomerase activity.

METHODSAccording to the coding sequence of hTERT mRNA, the target of RNAi was designed, and recombinant expression plasmid pshRNA-hTERT was constructed. The vector was transfected into Hep-2 cells. The radiosensitivity of Hep-2 cells was determined by clonogenic assay. Telomeric repeat amplification protocol (TRAP-PCR-ELISA) was used to observe the telomerase activity in each group. Results Recombinant expression vector pshRNA-hTERT was successfully transfected into Hep-2 cells. The hTERT expression inhibition rate reached 60. 8%. pshRNA-hTERT not only inhibited telomerase activity of Hep-2, but also inhibited the raise of telomerase activity induced by gamma-irradiation. Exposure of Hep-2 cells to pshRNA-hTERT for 24 hrs before irradiation resulted in a decrease in mean surviving fraction of Hep-2 cells compared with cells of group with irradiation alone (67. 7% vs 85. 7%, P <0. 05) .

CONCLUSIONRNAi showed a significant inhibitory effect to the expression of hTERT. The results indicate that pshRNA-hTERT can effectively inhibit telomerase activity of Hep-2 cells treated or untreated with 2 Gy gamma-irradiation and significantly enhance the radiosensitivity of Hep-2 cells in vitro. The role of radiosensitization of pshRNA-hTERT may be related with the inhibition of telomerase activity.

Carcinoma, Squamous Cell ; enzymology ; genetics ; pathology ; Cell Line, Tumor ; Cell Survival ; genetics ; radiation effects ; Cobalt Radioisotopes ; Enzyme-Linked Immunosorbent Assay ; Gamma Rays ; Humans ; Laryngeal Neoplasms ; enzymology ; genetics ; pathology ; Plasmids ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; biosynthesis ; Telomerase ; genetics ; metabolism ; Transfection

OBJECTIVE: To filter biomarkers of nasopharyngeal carcinoma (NPC) by constructing the homogenesis tissue gene expression profiling with the whole human genome GeneChip. METHODS: The epithelium cells of the homogenesis NPC and the pure nasopharyngeal normal tissues microdissected from nasopharyngeal biopsy which was preserved in the RNAlater were used to isolate RNA and then to harvest the aRNA through in vitro transcription, and aRNA prober was labled to hybridize to HG-U133. plus 2.0, so the expression profiling of each homogenesis tissue could be constructed. RESULTS: Some candidate biomarker genes related to the tumorigenesis of NPC had been filtered by comparing the expression profiling of NPC samples with the expression profiling of normal nasopharyngeal epithelia samples. Any genes regarding the metastasis of NPC might have been selected by comparing the expression profiling of no-metastasis samples with those of the metastasis samples. CONCLUSION Using the whole genome GeneChip to construct the expression profiling for the microdissected homogenesis tissue is effective to filter the candidate biomarker genes.
Adult ; Aged ; Biomarkers, Tumor ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Microdissection ; Middle Aged ; Nasopharyngeal Neoplasms ; genetics ; Nasopharynx ; metabolism ; Neoplasm Proteins ; biosynthesis ; genetics ; Oligonucleotide Array Sequence Analysis ; Tumor Suppressor Proteins ; biosynthesis ; genetics

BACKGROUNDThe NASG gene has been confirmed as a tumor-suppressor gene candidate related to nasopharyngeal carcinoma (NPC) by previous studies. We further investigated the expression and the role of NASG in the homogeneous tissue cells by microdissecting the samples of tissue from human NPC, and introduced a new way to study the expression of specific genes in tumor tissue.

METHODSThe RNAlater reagent was used to preserve the samples of tissue from the nasopharynx of NPC patients. The samples were microdissected to harvest the homogeneous tissue cells and then total RNA was isolated from them. The antisense RNA (aRNA) was amplified from the total RNA by "in vitro transcription (IVT)". We investigated NASG expression in the homogeneous tumor cells of NPC (22 samples) and compared it with that in the pure epithelial pillar cells of normal nasopharyngeal (10 samples) by semi-quantitative reverse transcription-polymerase chain reaction (sqRT-PCR).

RESULTSThe high quality total RNA could be harvested from the microdissected homogeneous tissue cells of the nasopharynx, then sufficient aRNA was derived from it. NASG gene expression was identified using aRNA by sqRT-PCR and showed that there was significant difference between the average value of case groups and that of control group (t = -5.275, df = 30, P < 0.001). The NASG gene in the subgroups WHOII tended to express lower levels than those in the subgroup WHOIII although this difference was not statistically significant (t = -1.584, df = 20, P = 0.129 > 0.05).

CONCLUSIONSMicrodissection was an effective method to obtain the homogeneous tissue cells of nasopharyngeal tissue (including the samples of NPC and non-NPC) in our study. Sufficient aRNA from amplifying total RNA could be used in sqRT-PCR to analyse the expression of NASG in the pure tissue cells. NASG should be a tumor-suppression gene candidate regarding to NPC.

Adult ; Aged ; Female ; Genes, Tumor Suppressor ; Humans ; Male ; Microdissection ; Middle Aged ; Nasopharyngeal Neoplasms ; genetics ; Nasopharynx ; metabolism ; RNA ; analysis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo analyze the clinical outcome of intracytoplasmic sperm injection (ICSI) in patients with previous fertilization failure after conventional IVF.

METHODSData from 20 ICSI cases (22 ICSI cycles) with previous complete failure of fertilization or with fertilization rate < or = 20% between January 2002 and December 2004 were retrospectively analyzed. The control group consisted of 100 consecutive ICSI cycles for male factor infertility in the same period.

RESULTSThe fertilization rate dramatically increased from 5.4% after conventional IVF to 76.9% after ICSI treatment (chi-squared = 264.66, P < 0.001). However, the fertilization rate in the subgroup with previous low fertilization was significantly lower than those in the control and in the subgroup without previous fertilization (67.9% vs 77.5%, 67.9% vs 84.2%). Compared with the control group, the subgroup without previous fertilization had a higher pregnancy rate and implantation rate, but only the difference in the implantation rate was statistically significant (40.5% vs 18.9%).

CONCLUSIONICSI can overcome previous fertilization failure with conventional in vitro fertilization and thus improve the clinical outcome.

Adult ; Case-Control Studies ; Female ; Fertilization in Vitro ; Humans ; Infertility ; therapy ; Male ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; Treatment Failure

OBJECTIVETo review the outcome of repeated percutaneous sperm aspiration (PESA) and testicular sperm extraction (TESE) for intracytoplasmic sperm injection (ICSI).

METHODSForty-three cycles of 31 cases of azoospermic patients which underwent at least two PESA or TESE for ICSI from January 2001 to December 2002 were collected. The sperm retrieval, fertilization, implantation and clinical pregnancy were analyzed.

RESULTSTwenty-four cases underwent PESA and 7 cases underwent TESE. There were not any complications in these patients. Compared with the first cycle of 154 cases, the fertilization rate, implantation rate and clinical pregnancy rate were 78.39% vs 73.64%, 19.68% vs 18.38% and 34.88% vs 37.91%, respectively(P > 0.05).

CONCLUSIONSRepeated PESA or TESE is safe and well tolerated in azoospermic patients. Compared with the first cycle, the differences of repeated PESA or TESE cycles in fertilization rate, implantation rate and clinical pregnancy rate were not statistically significant.

Adult ; Azoospermia ; therapy ; Female ; Humans ; Male ; Middle Aged ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic ; methods ; Tissue and Organ Harvesting ; methods

OBJECTIVETo develop a real-time fluorescent PCR protocol suitable for the routine screening of AZFc/DAZ microdeletions on the Y chromosome in azoospermic and oligozoospermic male infertility patients.

METHODSA set of real-time fluorescent PCR was established. Eighty-seven azoospermic and ligozoospermic patients undergoing ICSI in the IVF center and 30 azoospermic men undergoing testicular biopsy in the clinic of urology surgery were screened for AZFc/DAZ microdeletions of Y chromosome.

RESULTSEleven cases (9.4%) of AZFc/DAZ microdeletions were found in 117 cases of azoospermic and oligozoospermic patients by screening of realtime fluorescent PCR. Four cases (6.6%) were found in 61 oligozoospermic patients, and 7 cases (12.5%) were found in 56 azoospermic patients.

CONCLUSIONThe real-time fluorescent PCR protocol presented in this study is an easy and reliable method for detection of AZFc/DAZ microdeletions on the Y chromosome, which yields identical results to those of the multiplex PCR.

Chromosome Deletion ; Chromosomes, Human, Y ; Deleted in Azoospermia 1 Protein ; Fluorescence ; Humans ; Infertility, Male ; genetics ; Male ; Polymerase Chain Reaction ; methods ; RNA-Binding Proteins ; genetics

OBJECTIVETo evaluate the applicability of the polymorphic marker closely linked with beta-globin gene for the preimplantation genetic diagnosis (PGD) in couples at risk of having child with beta-thalassemia.

METHODSSingle cell multiplex nested PCR which coamplifies the beta-globin gene and the closely linked polymorphic marker, HumTHO1 gene, was applied in six clinical PGD cycles for four couples with beta-thalassemia.

RESULTSIn six clinical PGD cycles, a total of 44 embryos were biopsied and 44 blastomeres were obtained. Forty-one blastomeres were amplified and thirty-five embryos were given definite diagnoses. Fourteen embryos were transferred back to the uterus of the patients and one pregnancy went on well and ended with one live healthy birth, which confirmed the results of PGD. The average amplification efficiency of single blastomere was 89.7% and the average allele drop-out(ADO) rate was 14.4%. The coamplification of HumTHO1 could help to detect the existence of ADO and contamination.

CONCLUSIONThis is the first report on unaffected pregnancy resulting from PGD using multiplex nested PCR in China. The simultaneous amplification of polymorphic marker closely linked to beta-globin gene(HumTHO1) could help to resist the risk of misdiagnosis in PGD caused by ADO and contamination.

Adult ; Female ; Humans ; Male ; Polymerase Chain Reaction ; Pregnancy ; Preimplantation Diagnosis ; methods ; beta-Globins ; genetics ; beta-Thalassemia ; diagnosis ; genetics

OBJECTIVETo observe the effect on intervention of sub-health with pestle needle (Chuzhen).

METHODRandomized controlled trail was adopted for this research. One hundred and fifty-three cases were randomly divided into two groups of a Chuzhen group (79 cases) and a massage group (74 cases). Acupoint of Bazhen (Baihui Bazhen, Shendao Bazhen, Zhiyang Bazhen, Mingmen Bazhen, Yaoyangguan Bazhen), Hechelu on the head, the neck and the lumbar area were adopted in Chuzhen group. While regular whole-body massage was applied in the massage group. The human sub-health score, the cornell medical index (CMI) and thermal texture maps system (TTM) technology of the two groups before and after the intervention were observed.

RESULTS1) After treatment, sub-health condition score, the CMI score, the M-R score and the TTM index were all increased in both groups (all P<0.01) 2) Comparison of D-value of the two groups before and after the intervention: the level of the sub-health score, the total score of CMI, and the index of sleep, pressure, Governor Vessel, Hukou (first web), blood lipid, viscosity of blood, microcirculation of TTM index of the Chuzhen group changed more obvious (all P<0.01), but there was no statistic significances in the M-R score and blood sugar of the TTM (both P>0.05). 3) The sub-health condition score in Chuzhen group was higher than that in the massage group (P<0.01).

CONCLUSIONChuzhen therapy has definite effect on intervention of sub-health, which is better than regular general massage.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Cornell Medical Index ; Female ; Humans ; Male ; Massage ; Middle Aged ; Young Adult

OBJECTIVETo develop a multiplex PCR protocol, which could be suitable for routine screening of microdeletions on the Y chromosome in azoospermic and oligozoospermic male infertility patients.

METHODSFive multiplex sets were established. Eighty-seven azoospermic and oligozoospermic patients undergoing intracytoplasmic sperm injection (ICSI) in the in vitro fertilization (IVF) center and 30 azoospermic men undergoing testicular biopsy in the clinic of Urology Surgery were screened for microdeletions of Y chromosome.

RESULTSA total of 19 (16.2%) cases of microdeletions were found in 117 azoospermic and oligozoospermic patients by screening of Y chromosome microdeletions. Of these, 11 cases (18.0%) were found in 61 oligozoospermic patients, and 8 cases (14.3%) were found in 56 azoospermic patients.

CONCLUSIONThe multiplex PCR protocol presented in this study is an easy-to-do and reliable method for detecting microdeletions on the Y chromosome. Routine screening of microdeletions on the Y chromosome for azoospermic and oligozoospermic patients is essential.

Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Female ; Genetic Testing ; methods ; Humans ; Infertility, Male ; diagnosis ; genetics ; Male ; Polymerase Chain Reaction