Objective:To establish the ISSR fingerprint of Chrysanthemum morifolium cultivated in Futianhe area of Macheng county to guide the breeding of C. morifolium. Methods:Using the technology of ISSR molecular markers and the software of SPSS 15. 0, the coefficient matrix of Jaccard was established and the tree graph of the genetic relationship of the breeds of C. morifolium cultivated in Fu-tianhe area was built to analyze the respective genetic relationship and features. Results:By ISSR analysis, it confirmed that C. morifoli-um in Futianhe area had long genetic distance with the other white chrysanthemum breeds, and it could be considered as an individual breed. Conclusion:The ISSR map can be used to identify the breed of C. morifolium cultivated in Futianhe area.

Objective To present our initial experience of using transumbilical multi-stab laparoscopic pyeloplasty(TMLP) as a treatment for the infants younger than 3 months with severe hydronephrosis caused by ureteropelvic junction obstruction (UPJO).Methods Seventy-two infants younger than 3 months with severe hydronephrosis caused by UPJO underwent TMLP from June 2010 to March 2013 in our center.The average age received operation was 34 days (7-87 days).The operative indications included:① prenatal diagnosis of Society of Fetal Urology Grade 4 hydronephrosis; ②ipsilateral differential renal function being less than 40％.Patients were followed up with physical examinations,ultrasound and radionuclide scans.Results The operations were successfully performed in all the 72 patients.Neither conversion nor additional trocars placement was required and there was no intraoperative complication.The average operative time was 75 min (range,53-118 min).The patients were followed up for 6-36 months (mean,12 months).The renal pelvic anteroposterior diameters were reduced and the renal functions were improved (P＜ 0.05).The anastomoses were proved to be patent and the renal parenchymal thickeness increased.In addition,the scars were barely noticeable.Conclusions TMLP for the infants younger than 3 months with severe hydronephrosis caused by UPJO is feasible,safe and less invasive.The cosmetic results are excellent.But the surgery must be operated by rich experienced surgeon.

Objective To summarize the clinical features of 22 probands diagnosed with congenital muscular dystrophy (CMD),and to provide genetic counseling and prenatal diagnosis for 23 fetuses of these pedigrees.Methods Data of 22 CMD patients who were treated in the Pediatric Department of Peking University First Hospital during October 2006 to March 2016 were analyzed.Informed written consents for participation in this study were obtained from the parents or guardians.Prenatal diagnosis was performed using DNA samples extracted from fetal villus cells of 12 cases at 11-13 gestational weeks and amniotic fluid of 11 cases at 18-22 gestational weeks.Direct DNA sequencing by polymerase chain reaction (PCR) and multiplex ligation-dependent probe amplification (MLPA) were used to detect CMD-related gene mutations.Linkage analysis of short tandem repeats (STRs) was used to identify maternal blood contamination and biological parents.Results Thirteen out of the 22 probands with CMD were diagnosed with congenital muscular dystrophy type 1 A (MDC1A),and all of them carried compound heterozygous mutations in LAMA2 gene.Prenatal diagnosis of 13 fetuses from these pedigrees found that four fetuses were wild-type,seven were heterozygotes and two carried the same mutations as their proband.Three probands with LMNA-related congenital muscular dystrophy (L-CMD) carried de novo mutations in LMNA gene.In these pedigrees,two fetuses were wild-type and one whose mother was mosaicism carried the same mutations as the proband.One proband with Ullrich congenital muscular dystrophy carried compound heterozygous mutations in COL6A2 gene and the fetus of the same pedigree was wild-type.Five probands were diagnosed with α-dystroglycanopathies.And among them,two cases of muscle-eye-brain disease (MEB) carried compound heterozygous mutations in POMGnT1 gene and the fetuses of the two peidgrees were heterozygotes;one case of congenital muscular dystrophy type 1C (MDC1C) had compound heterozygous mutations in FKRP gene and the fetus carried the same mutations;one patient diagnosed with POMGnT1-related congenital muscular dystrophy with mental retardation (CMD-MR) carried compound heterozygous mutations in POMGnT1 gene,and the fetus was positive for the same mutations;one proband with POMT1-related CMD-MR was positive for compound heterozygous mutations in POMT1 gene and the results of prenatal diagnosis for two fetuses of this pedigree showed that the first fetus had the same mutations as the proband,while the second was heterozygote.Conclusions No effective therapeutic method is available for CMD.Therefore,accurate genetic counseling and prenatal diagnosis are necessary to prevent CMD child from birth.

Objective To construct the bait plasmid of HNP-3 mature peptide in yeast two-hybrid system and examine whether the recombinant bait plasmid has self-activating and toxicity effect.Methods Using RT-PCR technique,the cDNA fragments of HNP-3 mature peptide gene were amplified from the extracted RNA in cultured HL-60 cells.The fragment was firstly cloned into pBluescript-SK-II vector,confirmed by sequencing,then sub-cloned into the bait plasmid pGBKT7 and identified with PCR and sequence analysis techniques.The recombinant plasmid was introduced into the yeast cell AH109,and its self-activating and toxicity effect was tested by auxotrophic selective culture.Results DNA sequencing indicated that the inserted fragment in pBluescript-SK-II vector was HNP-3 mature peptide gene sequence,and the sub-cloned recombinant pGBKT7-HNP-3 was no mismatch.The recombinant bait plasmid didn't have self-activating effect and did not show toxicity to yeast AH109 cell.Conclusion The bait plasmid of HNP-3 mature peptide was constructed successfully.This was helpful for investigating the proteins interacting with HNP-3 mature peptide by yeast two-hybrid technique.

Objective To summarize the experience of patient positing,port placements setting and robot cart docking in pediatric robot-assisted laparoscopic upper urinary tract operations.Methods From March 2017 to December 2017,140 robot-assisted laparoscopic upper urinary tract procedures were performed in our institution,including 110 cases of pyeloplasty,15 upper pole heminephroureterectomy,12 simple nephrectomy and 3 adrenalectomy.There were 103 males and 37 females with a range age from 1 month to 18 years.The assistant surgeon was adjacent to the instrument nurse,and patients were placed in a supine position with 60°-80° inclination and keep the legs low to the body.Room setup and patient positioning were similar to the traditional laparoscopic surgery.Semi-hidden incision technique was used in 140 patients:the camera port was placed umbilicus,two additional arm ports (one 5 mm and one 8 mm) were placed under direct vision,the 8 mm arm port was placed on the line of a Pfannenstiel incision and the 5 mm arm port was placed below the Xiphoid along the midline.Finally,a 3 or 5 mm assistant port was placed approximately 3 cm lateral to the inferior arm port,the line of a Pfannenstiel incision.Results The average time was (11.5 ± 3.2) min (10.5-16.5 min) from skin incision to robot cart docking completed.All surgeries were successfully completed without open conversion.One patient required an additional assist port for severe adhesion after the previously open surgery,there was no injury to other viscera.Average operative time was (146.9 ± 48.7)min (78-259 min) and average post-operative hospitalization time was (5.7 ± 1.4) d(4-10 d),respectively.There was no visual scar on abdominal 6 weeks postoperatively,and all parents made comments about their satisfaction with the cosmetic appearance.All operations got complete success at a mean follow up of 6 (1-9) months.Conclusions A good room setup,patient positioning and the semi-hidden incision technique port placements are maintaining the safety of the patient,avoiding compression injuries,allowing maximum mobility of the robotic arms,and facilitating a smooth and efficient surgery,and improving post-operative recovery.

BACKGROUND:Ischemic postconditioning is one of the effective ways to reduce ischemia-reperfusion injury and has been more and more widely used in clinical practice in recent years,but its specific molecular mechanism has yet to be studied. OBJECTIVE:To investigate the role and mechanism of piRNA-005854 in the aging cardiomyocytes caused by hypoxic postconditioning. METHODS:In vitro,cardiomyocytes were administered 8 mg/mL D-galactose for 9 days to induce their aging.β-Galactosidase staining was used to observe the aging of cardiomyocytes.Senescent cells were treated with hypoxia/reoxygenation and hypoxic postconditioning.ELISA was utilized to detect changes in myocardial injury markers creatine kinase isoenzyme MB and lactate dehydrogenase levels.Western blot assay was applied to detect the expression changes of autophagy-related proteins LC3II,p62,ULK1 and phosphorylated ULK1 in aging cardiomyocytes.qRT-PCR was employed to determine the expression level of piRNA-005854.piRNA-005854 inhibitor and piRNA-005854 mimics were transferred into aging cardiomyocytes and followed with hypoxic postconditioning.Western blot assay was used to examine the expression of LC3II,p62,ULK1 and phosphorylated ULK1. RESULTS AND CONCLUSION:(1)D-galactose induced obvious senescence of cardiomyocytes 9 days later.(2)Compared with the normoxia group,creatine kinase isoenzyme MB and lactate dehydrogenase levels increased in the hypoxia/reoxygenation group(P<0.01);LC3 II/I expression was increased;p62 expression was decreased;ULK1 phosphorylation level was increased,and piRNA-005854 expression was increased(P<0.01).(3)Compared with the hypoxia/reoxygenation group,creatine kinase isoenzyme MB and lactate dehydrogenase levels significantly reduced in the hypoxic postconditioning group(P<0.01);LC3 II/I expression significantly decreased(P<0.05);p62 expression increased(P<0.01);ULK1 phosphorylation level decreased(P<0.05),and piRNA-005854 expression decreased(P<0.01).(4)After transfection of piRNA-005854 inhibitor,LC3II/I expression was decreased(P<0.01);the expression of p62 was increased significantly(P<0.05);the phosphorylation level of ULK1 was decreased significantly(P<0.01).After transfection of piRNA-005854 mimics,LC3II/I expression was increased significantly;the expression of p62 was decreased,and the phosphorylation level of ULK1 was increased significantly(P<0.01).(5)The results show that piRNA-005854-mediated reduction of ULK1-dependent autophagy level is a possible mechanism that hypoxic postconditioning exerts its protective effect on aging cardiomyocytes.

Background Exposure to polycyclic aromatic hydrocarbons (PAHs) is associated with the development of metabolic syndrome (MS). However, the role of amino acids in PAH-induced MS remains unclear. Objective To explore the impact of PAHs exposure on the incidence of MS among coking workers, and to determine potential modifying effect of amino acid on this relationship. Methods Unmatched nested case-control design was adopted and the baseline surveys of coking workers were conducted in two plants in Taiyuan in 2017 and 2019, followed by a 4-year follow-up. The cohort comprised 667 coking workers. A total of 362 participants were included in the study, with 84 newly diagnosed cases of MS identified as the case group and 278 as the control group. Urinary levels of 11 PAH metabolites and plasma levels of 17 amino acids were measured by ultrasensitive performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Logistic regression was used to estimate the association between individual PAH metabolites and MS. Stratified by the median concentration of amino acids, Bayesian kernel machine regression (BKMR) model was employed to assess the mixed effects of PAHs on MS. Due to the skewed data distribution, all PAH metabolites and amino acids in the analysis were converted by natural logarithm ln (expressed as lnv). Results The median age of the 362 participants was 37 years, and 83.2% were male. Compared to the control group, the case group exhibited higher concentrations of urinary 2-hydroxyphenanthrene (2-OHPhe), 9-hydroxyphenanthrene (9-OHPhe), and hydroxyphenanthrene (OHPhe) (P=0.005, P=0.049, and P=0.004, respectively), as well as elevated levels of plasma branched chain amino acid (BCAA) and aromatic amino acid (AAA) (P<0.05). After being adjusted for confounding factors, for every unit increase in lnv_2-OHPhe in urine, the OR (95%CI) of MS was 1.57 (1.11, 2.26), and for every unit increase in lnv_OHPhe, the OR (95%CI) of MS was 1.82 (1.16, 2.90). Tyrosine, leucine, and AAA all presented a significant nonlinear correlation with MS. At low levels, tyrosine, leucine, and AAA did not significantly increase the risk of MS, but at high levels, they increased the risk of MS. In the low amino acid concentration group, as well as in the low BCAA and low AAA concentration groups, it was found that compared to the PAH metabolite levels at the 50th percentile (P₅₀), the log-odds of MS when the PAH metabolite levels was at the 75th percentile (P₇₅) were 0.158 (95%CI: 0.150, 0.166), 0.218 (95%CI: 0.209, 0.227), and 0.262 (95% CI: 0.241, 0.282), respectively, However, no correlation between PAHs and MS was found in the high amino acid concentration group. Conclusion Amino acids modify the effect of PAHs exposure on the incidence of MS. In individuals with low plasma amino acid levels, the risk of developing MS increases with higher concentrations of mixed PAH exposure. This effect is partly due to the low concentrations of BCAA and AAA.