AIM AND METHODS: Using Ca 2+ -sensitive fluorescent probe Fura-2,we measured the changes of _i in cultured rat pulmonary artery endothelial cells (PAEC) and porcine pulmonary artery smooth muscle cells (PASMC) from normoxic (NC group) or chronic hypoxic group (CH group) when they were exposed to acute hypoxia. RESULTS: The increase in _i in 6th passage of PASMC caused by acute hypoxia in CH group was significantly lower than that in the same passage of NC group (P

BACKGROUND:The main clinical manifestation of senile arteriosclerosis obliterans is lower limb ischemia, which is currently difficult to treat. One method is by autologous stem cell transplantation into the muscles of ischemic limbs to improve the formation of new capillaries and restore lower limb blood flow. Endothelial progenitor cell marker CD34+ cell transplantation has been shown to promote angiogenesis in ischemic limbs. Therefore, we propose that peripheral blood autologous CD34+ cell transplantation in older adult patients with atherosclerotic ischemia could effectively promote angiogenesis.OBJECTIVE:To assume that peripheral blood autologous CD34+ cell transplantation in the elderly with atherosclerotic ischemia could effectively promote angiogenesis.METHODS:This is a prospective, single-center, open-label, randomized, and controlled clinical trial that will be completed at the Qingdao No. 9 People's Hospital, China. Twenty older adult patients with atherosclerotic lower limb ischemia will be randomized into two groups. In the cell transplantation group (n=10), peripheral blood CD34+ cells transfected with vascular endothelial growth factor 165 (VEGF165) gene will be intramuscularly transplanted into the ischemic limbs in older adult patients with atherosclerotic lower limb ischemia. In the control group (n=10), normal saline will be intramuscularly injected into the ischemic limbs. All patients will be followed up for 6 months. The primary outcome will be ankle-brachial indices before and 6 months after transplantation to assess lower limb ischemia in both groups.The secondary outcomes will be the number of microvessels in the lower limb muscles before and 6 months after transplantation, the morphology of new blood vessels revealed by CT angiography, the number of VEGF-immunoreactive cells 6 months after transplantation and the incidence of adverse reactions. The trial was registered at the ClinicalTrials.gov (identifier:NCT03098771), and the study protocol was approved by the Ethics Committee of Qingdao No. 9 People's Hospital of China. All protocols will be in accordance with Declaration of Helsinki,formulated by the World Medical Association. All patients will be informed of study protocols and provide a written informed consent prior to the beginning of the trial.DISCUSSION:This trial will begin in January 2018 and finish in December 2019. We aim to quantify the effects of VEGF165 gene-modified CD34+ cell transplantation in the treatment of older adult patients with atherosclerotic ischemia to develop a new effective treatment of lower limb ischemia.

Objective　To construct rapid amplification cDNA ends(RACE) cDNA libraries from human fetal bone and joint and provide resources for isolation of bone- and joint- specific development-related genes.Methods　Total RNA of bone and joint were extracted with the modified single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. The double-stranded end-blunted cDNA were synthesized using TaKaRa's cDNA synthesis kit and ligated to cassette adaptors. All of the cDNA molecules were amplified by a pair of common primers.Results　A protocol for RACE cDNA library construction from bone and joint was established and two RACE cDNA libraries from human fetal bone and joint were successfully constructed.Conclusion　The protocol of RACE cDNA library construction from limited materials proved to be simple and efficient and the library was suitable for RACE to isolate tissue-specific genes.

Preeclampsia is one of the main causes of high morbidity and mortality of pregnant women and perinatal infants worldwide. Affected by many factors, preeclampsia has a complex pathogen-esis and can cause involvement of multiple organs and systems. Its pathogenesis is still unclear. Current research suggests that maternal immune system indirectly involved in the pathophysiology of preeclampsia change, that is, abnormal activation of innate immune cells and unbalanced differentiation of T helper cell subsets interfere with normal immune regulation, and interact with inflammatory response of the body, which produces cytotoxic environment at the maternal-fetal interface and affects trophoblast inva-sion. Therefore, clarifying the role of the immune system can not only clarify the pathogenesis of pre-eclampsia, but also contribute to the development of diagnosis and treatment of preeclampsia. This paper reviews the research status of immune system in preeclampsia, including innate immunity and adaptive immunity . The immune mechanism of preeclampsia is elaborated mainly from immune regulation mediated by T lymphocyte, natural killer ( NK) cell, macrophage and human leukocyte antigen.

Objective:The correlation between in vitro fertilization-embryo transfer (IVF-ET) pregnancy and preeclampsia was studied by the propensity score matching. Methods:4 823 pregnant women with delivery gestational weeks >24 weeks were selected, including 481 in IVF group and 4 342 in natural pregnancy group. The propensity score model was established by using 16 maternal covariates, and the propensity score matching samples (924 cases) were obtained to evaluate the correlation between IVF-ET and preeclampsia.Results:⑴ Before matching, the incidence of preeclampsia in the IVF group was higher than that in the natural pregnancy group (9.8% vs 3.3%, P<0.05). Multivariate regression analysis showed that IVF-ET was a risk factor for preeclampsia (a OR=1.887; 95% CI: 1.23-2.89, P=0.003); After matching propensity score, OR was 2.067 (95% CI: 1.24-3.44, P=0.005), confirming that there was a significant association between IVF-ET and preeclampsia. ⑵ Before matching, the incidence of preeclampsia in IVF group was significantly higher than that in natural pregnancy group in singleton pregnancy (9.0% vs 3.1%, a OR=2.530, 95% CI: 1.63-3.94, P>0.05); In twin pregnancy, there was no significant difference in the incidence of preeclampsia between the two groups (12.7% vs 7.5%, a OR=1.004, 95% CI: 0.35-2.87, P=0.994); The result of propensity score matching is consistent with that before matching. Conclusions:Propensity score matching analysis showed that the risk of preeclampsia increased after IVF-ET pregnancy, IVF-ET was an important risk factor for preeclampsia in singleton pregnancy, and IVF-ET did not increase the risk of preeclampsia in twin pregnancy. It is suggested that the correlation between IVF and preeclampsia may be disturbed by twin pregnancy.

Objective　To analyze the genetic polymorphism of 8 STR loci on chromosome 11 and 5 STR loci on chromosome 19 in Chinese Hans. Methods　Polymerase chain reaction-single strand length polymorphism(PCR-SSLP) was used to genotype 100 randomly selected individuals of the Han nationality at 8 STR loci(D11S1984, D11S1999, D11S1392, D11S1985, D11S2002, D11S1986, D11S4464 and D11S2359) on chromosome 11 and 5 STR loci(D19S247, D19S714, D19S433, D19S246, D19S254) on chromosome 19. Results　Eight alleles and 26 genotypes, 9 alleles and 15 genotypes, 6 alleles and 16 genotypes, 12 alleles and 40 genotypes, 6 alleles and 19 genotypes, 12 alleles and 48 genotypes, 8 alleles and 20 genotypes, 7 alleles and 13 genotypes were observed at D11S1984, D11S1999, D11S1392, D11S1985, D11S2002, D11S1986, D11S4464 and D11S2359. The heterozygosities for the 8 STR loci were 87%, 68%, 73%, 92%, 71%, 86%, 75% and 71%, respectively. Ten alleles and 19 genotypes, 10 alleles and 26 genotypes, 10 alleles and 24 genotypes, 11 alleles and 29 genotypes, 8 alleles and 18 genotypes were observed at D19S247, D19S714, D19S433, D19S246, D19S254. The heterozygosities for the 5 STR loci were 63%, 82% 72%, 81% 74%, respectively. Conclusion　 The distribution of allele frequencies of 8 STR loci on chromosome 11 and 5 STR loci on chromosome 19 was consistent with the Hardy-Weinberg equilibrium and the highly genetic polymorphism was observed in Chinese Han population.

OBJECTIVETo investigate the relationship between polymorphism of N-acetyltransferase (NAT2) gene and genetic susceptibility to laryngeal carcinoma.

METHODSA case-control study on 62 laryngeal carcinoma patients and 56 controls was conducted. NAT2 alleles were differentiated by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) methods using originally created PCR primers and genomic DNA extracted from peripheral white blood cells. Genetic risk for NAT2 genotype was analyzed by smoking index (SI, cigarettes smoked per day x years of smoking).

RESULTSThe frequency of NAT2 slow genotype was 80.6% in patients with laryngeal carcinoma and 60.7% in the controls, the difference of which was statistically significant (chi(2) = 5.70, P = 0.017). The odds ratios were 2.70 (95% CI 1.19 approximately 6.11). Among the individuals with NAT2 slow genotype at high level of cigarette smoking, there was a significantly higher risk of 5.64 (95% CI 1.77 approximately 17.92), while those at low level were considered the reference group (OR 1.38, 95% CI 0.42 approximately 4.52).

CONCLUSIONNAT2 slow genotype increases the risk of susceptibility to laryngeal carcinoma. The combined effect of NAT2 slow genotype and exposure to smoking is observed during the development of laryngeal cancer.

Aged ; Alleles ; Arylamine N-Acetyltransferase ; genetics ; DNA, Neoplasm ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Polymorphism, Genetic ; Smoking ; Statistics as Topic

OBJECTIVETo determine the linkage between Smith-Fineman-Myers syndrome (SFMS) and X-linked nuclear protein(XNP) locus.

METHODSPolymerase chain reaction and denaturing polyacrylamide gel electrophoresis were used to genotype two polymorphic short tandem repeats within XNP gene.

RESULTSOne of the two short tandem repeats was informative in SFMS family from Shandong, China. Recombination between SFMS locus and XNP gene was observed in the SFMS family.

CONCLUSIONXNP gene is not associated with the disease in the SFMS family from Shandong, China. SFMS exhibits locus heterogeneity at molecular level.

Abnormalities, Multiple ; genetics ; Craniofacial Abnormalities ; genetics ; DNA Helicases ; Female ; Genetic Linkage ; Growth Disorders ; genetics ; Humans ; Intellectual Disability ; genetics ; Male ; Muscle Hypotonia ; genetics ; Nuclear Proteins ; genetics ; Pedigree ; Phenotype ; Polymorphism, Genetic ; Recombination, Genetic ; Syndrome ; X Chromosome ; X-linked Nuclear Protein

OBJECTIVETo determine the genomic structure of low density lipoprotein receptor related protein 5 (LRP5) gene.

METHODScDNA sequence encoding LRP5 was used to screen genomic clones containing LRP5 gene by computer hybridization approach. By comparing the cDNA sequence of LRP5 with the genomic sequences, the genomic structure of LRP5 was determined, and then it was conformed by amplifying and sequencing the sequences of exons and splicing junction.

RESULTSThe genomic sequence of LRP5 gene was 131.6 kb in length, containing 23 exons and 22 introns. Three single nucleotide polymorphisms were detected within the coding sequences of LRP5 gene, namely A459G in exon 2, C2220T in exon 10 and G4416C in exon 21. Four polymorphic markers, D11S1917, D11S4087, D11S1337 and D11S4178, located in the 5' flank sequence, introns 1, 4, and 13 of the LRP5 gene, respectively.

CONCLUSIONThe characterization of genomic structure of LRP5 gene allows the investigators to detect disease-causing mutation within the gene and further study the function of LRP5 gene.

Base Sequence ; DNA ; chemistry ; genetics ; Exons ; Genes ; genetics ; Humans ; Introns ; LDL-Receptor Related Proteins ; Low Density Lipoprotein Receptor-Related Protein-5 ; Polymorphism, Single Nucleotide ; Receptors, LDL ; genetics ; Sequence Analysis, DNA

Objective: To evaluate cardiopulmonary exercise testing (CPET) on sildenaifl effect for treating the patients with pulmonary arterial hypertension (PAH). Methods: A total of 25 PAH patients received sildenaifl treatment in our hospital from 2012-01 to 2014-01 were enrolled as PAH group, in addition, there were a Control group including 24 healthy subjects. The CPET, echocardiography, NYHA function class, 6-mimute walking distance (6MWD) and plasma levels of NT-proBNP at the baseline, (6-12) months and (13-18) months after sildenaifl treatment were assessed and compared between 2 groups. Results: Compared with Control group, PAH group showed decreased aerobic capacity (peakVO?2, Peak O2pulse) and ventilation efifciency (PETCO2@AT, VE?/VC?O2@AT), allP<0.05. At (8±2) months after sildenaifl treatment, aerobic capacity and ventilation efifciency was improved, meanwhile, NYHA function class, 6MWD and plasma levels of NT-proBNP were improved, allP<0.05. At (16±2) months after sildenaifl treatment, 6MWD was similar,P=0.26, while peak VO?2 and peak O2 pulse were decreased than they were at (8±2) months after sildenaifl treatment,P=0.04 and 0.06; the ventilation efifciency was elevated (as presented by increased VE?/VC?O2@AT and decreased PETCO2@AT,P=0.04 and P=0.04); plasma level of NT-proBNP was increased,P=0.05. Conclusion: CPET can effectively evaluate sildenaifl effect for treating PAH patients and therefore and guide the drugs therapy.