Objective To isolate the microsatellite DNA of Phlebotomus chinensis and screen the polymorphic loci.Methods Genomic DNA fragments were hybridized with biotinylated oligonucleotide probes(AAT)17,(GA)25,(CCT)17,and(TG)18.The hybridized fragments were captured with Vectrex Avidin D and enriched by centrifugal ultrafiltration with ultra-4-column ultrafiltrate.The target fragments were amplified,cloned and sequenced.The suitable microsatellite loci were chosen in Ph.chinensis's library to establish PCR amplification assay.The polymorphism screening was conducted by PAGE with Ph.chinensis field specimens.Results The enrichment protocol used in this study was efficient,with a percentage of recombinant clone as 78.6%.There were 118 microsatellite sequences in library,the GenBank accession numbers were from FJ919812 to FJ919932(except GenBank accession numbers FJ919833,FJ919836,and FJ919869).There were 72 typical microsatellite sequences occupying 61.0% and the rest were 46 nontypical microsatellite sequences in the library.Twenty-two loci were chosen to polymorphism screening and PAGE showed that 14 loci were polymorphic.The loci of dinucleotide repeat were more polymorphic than those of trinucleotide and polynucleotide repeat.Conclusion The microsatellite-containing library of Ph.chinensis has been constructed with 118 sequences,and 14 new polymorphic microsatellite loci are reported.

Purpose To explore the relationship of Tei index and the concentration of plasma brain natriuretic peptide (BNP) in patients with dilated cardiomyopathy. Materials and Methods Fifty healthy individuals (control group) and fifty dilated cardiomyopathy patients (study group) were recruited in this study. BNP level was measured by enzyme-linked immunosorbent assay (ELISA). Color Doppler echocardiography were performed to measure the cardiac indices including left ventricular end-diastolic dimension (LVEDd), left ventricular end-systolic diameter (LVESd), interventricular septal thickness at diastole (IVSd), left ventricular ejection fraction (LVEF). Tei index was then calculated. The indices between the two groups were compared and correlation of Tei index and BNP with cardiac indices was analyzed. Results LVEDd, LVESd, Tei index, and BNP were significantly higher in study group than those in control group (t=14.4, 23.4, 21.9 and 22.3, P<0.01). IVSd and LVEF were significant lower in the study group (t=12.4 and 12.5, P<0.01). BNP was negatively correlated with LVEF (r= -0.266, P<0.05), and positively correlated with Tei index and LVEDd (r=0.401 and 0.326, P<0.05). Tei index was negatively correlated with LVEF and E/A (r= -0.480 and - 0.241, P<0.05), and positively correlated with LVEDd (r=0.375, P<0.05). Multiple stepwise regression analysis showed that the variates of BNP stepwise regression equation were Tei index, LVEF and LVEDd (t=5.984, -2.477 and 2.326, P<0.05); after correcting LVEF, the stepwise regression analysis showed closer relationship between Tei index and BNP (t=2.728, P<0.05). Conclusion There is correlation between Tei index and BNP. Tei index may be a good index for accurately estimating global ventricular function.

Objective To investigate the effects of antisense p21 oligodeoxynucleotide (p21 ASODN) on the expression of p21 protein and extracellular matrix in cultured human glomerular mesangial cells (HGMC) under high glucose medium. Methods HGMCs were transfected with 50 nmol/L or 100 nmol/L p21 ASODN or scrambled control oligodeoxynucleotide (SCODN) using lipofectamine 2000. After incubation under normal (5.5 mmol/L) or high (30 mmol/L) glucose media and different times, HGMCs lysates were collected and the expression of p21, fibronectin and laminin was examined by Western blot. The secretion of fibronectin and laminin by HGMCs in supernatants of culture media was also detected with EOSA. Results High glucose media significantly stimulated the expression of p21, increased the syntheses and secretion of fibronectin and laminin in cultured HGMCs in a time-dependent manner. Transfection of HGMCs with p21 ASODN not only decreased p21 protein level caused by high glucose media, but also attenuated the expression and secretion of fibronectin and laminin in supematants of HGMCs lysates and culture media. Meanwhile, SCODN had no significant effects on the expression of p21, fibronectin and laminin. Conclusions High glucose promotes the expression of p21 , fibronectin and laminin in cultured HGMCs. Transfection of HGMCs with p21 ASODN can decrease p21 protein level caused by high glucose media, and attenuate the expression of fibronectin and laminin in supematants of HGMCs lysates and culture media. Modulation of p21 expression in HGMCs may work as an effective way to mitigate the progression of diabetic nephropathy.

ssay provides a higher detection rate for HCV infection in HD patients.

Objective To study conventional laparoscopic instruments line single hole peritoneoscope gallbladder excision and three hole laparoscopic cholecystectomy surgery efficacy and safety.Methods 140 cases patients with gallbladder stones in our hospital from January 2014 to June 2015 were selected as the research subjects.All patients need to be treated with cholecystectomy.All the patients according to the random number table were randomly divided into two groups,respectively for single hole group and three group.Single hole group using a single hole peritoneoscope gallbladder excision,triplex group use three hole laparoscopic gallbladder resection.The surgical effect,hospitalization costs and complications were compared between the two groups after treatment.Results Hole group operation time (87.89 ± 12.81) min,longer than the three-hole group (53.89 ± 8.91) min,but the hospital stay was (1.28 ± 0.21) d and total hospital costs (11 241.21 ± 23.91) yuan,were lower than three-hole group,P ＜ 0.05,the difference was statistical significance in the amount of bleeding in the two groups were not statistical significance,P ＞ 0.05;two groups of patients had complicatiom,but have been treated better,and two concurrent disease (1.43％ vs 2.86％) incidence was no significant difference,P ＞ 0.05.Conclusion Single hole laparoscopic cholecystectomy reduce the hospitalization time and hospitalization expenses,surgical trauma is smaller,postoperative recovery is faster,and the safety is high,it is worthy of clinical application.

Nine hundred and forty nine general physicians in Beijing urban area received surgical skill assessment in 2008.The assessment results of five surgical skills (taking out stitches,changing dressings,skin suture,knot with gloves and wearing isolation gown) were analyzed with the relations of different gender,age,educational background,professional title,specialty,hospital grade,length of service and times of test taking.The average score of the total assessment was 71.The score was lower in the practitioners who were male,older than 50 years,higher educational background,lower professional tide,more than 2 times of test taking,and who were specialized in traditional Chinese medicine,ophthalmology and otorhinolaryngology and emergency medicine.Results suggest that the levels of surgical skills in general practitioners are generally low and periodical training is needed.

Liquid biopsy is a new noninvasive detection method and is also the main molecular detection method to guide the application of precision medicine. Liquid biopsy mainly includes the measurement of circulating free DNA (cf-DNA), circulating tumor cells, and exosomes, and in particular, cf-DNA is becoming a valuable molecular detection tool, especially in the early detection and diagnosis of liver cancer and parasitic infections involving the liver. This article reviews the clinical application of cf-DNA in liver cancer and liver-related parasitic diseases.

objective To investigate the effects of different sections of receptor associated protein (RAP) on the expression and distribution of TRPC6,synaptopodin and podocalyxin in passive Heymann nephritis(PHN). Methods Male Sprague-Dawley rats were injected with three kinds of antisera (anti-RAP full-length serum,anti-RAP N-terminal serum and anti-RAP C-terminal serum)to establish three kinds of PHN models.The control group was injected with normal rabbit serum.The quatitation of 24 h urinary protein,serum albumin and creatinine were taken before injection and one week after PHN model successfully induced.The histopathologic changes of renal tissues were observed by light microscopy.The expression and distribution of TRPC6,synaptopodin and podocalyxin in glomerular podocytes were observed by laser scanning confocal microscopy and analyzed by fluorescence quantitative software after indirect immunofluorescence double staining.Results The quantities of 24 h urinary protein in the three model groups were significantly higher than those of themselves before injection and control groups (P<0.01,respectively).The values of serum albumin and creatinine were not significantly different before and after injection (P>0.05).The expression of TRPC6 in podocytes was higher in the PHN model groups than that of control group.Fluorescence intensity of TRPC6 in RAP full-length group was stronger than that in RAP N-terminal or C-terminal groups.The expressions of synaptopodin and podocalyxin distributed along the glomerular basement membrane as spot,discontinuous short line and defect of some segments,and were lower in three PHN groups than those of control group.Fluorescence intensity of synaptopodin and podocalyxin among three PHN groups had no differences. Conclusions RAP full-length and N-terminal or C-terminal parts can increase the expression of podocyte TRPC6,but decrease the expressions of synaptopodin and podocalyxin,and alter their distribution,which may be associated with the proteinuria,however,their role in the PHN pathogenesis needs further study.

ObjectiveTo evaluate the clinical value of prenatal ultrasonography combined with maternal serology screening for chromosomal abnormality in 15 to 20+6 gestational weeks.MethodsSix hundred and twenty-eight pregnant women (628 fetuses) in 15 to 20+6 gestational weeks were selected to undergo prenatal ultrasonography, who were in critical risk of trisomy 21 or trisomy 18 by maternal serology screening. Transabdominal ultrasonography were performed and fetal nasal bone and nuchal fold were detected. Those who had nasal bone hypoplasia, thickened nuchal fold (NF>6 mm) and other abnormal fetal soft markers underwent amniocentesis for karyotyping analysis. ResultsThere were 6 cases of nasal bone hypoplasia (0.96%, 6/628), including one case of thickened nuchal fold, two cases of echogenic bowel, two cases of choroid plexus cysts and 1 case of echogenic cardiac focus. All these 6 cases underwent amniocentesis and 2 were trisomy 21 (33.3%, 2/6). The other 4 cases had no significant chromosomal abnormality.ConclusionsPrenatal ultrasonography may improve the detection rate of chromosomal abnormality for those pregnant women who are in critical risk of chromosomal abnormality prompted by serology screening. But invasive procedures are still needed to verify the chromosomal abnormality.

Objective: To explore the antagonistic effect of quercetin on fine particulate matter (PM2.5)-induced embryonic developmental toxicity in vitro.Methods: PM2.5 was collected on glass fiber filters by PM2.5 samplers during the heating period of Dec.2015 to Mar.2016 in an area of Haidian District, Beijing City.The sampled filters were cut into 1 cm×3 cm pieces followed by sonication.The PM2.5 suspension was filtered into a 10 cm glass dish through 8 layers of sterile carbasus and stored at-80 ℃ until freeze drying.Frozen PM2.5 suspension was dried by vacuum freeze-drying.In vitro post-implantation whole embryo culture was used in this study.Pregnant rats with 9.5 gestation days (GD) were killed by cervical dislocation and the uteri were removed into sterile Hank's solution.The embryos with intact yolk sacs and ecto placental cones were induced by PM2.5, and then subjected to intervention of quercetin at the doses of 0.1 μmol/L, 0.5 μmol/L, 1.0 μmol/L and 5.0 μmol/L, respectively.At the end of the 48 h culture period, the cultures were terminated, and all embryos were removed from the culture bottles and placed in prewarmed Hank's solution for evaluation.Morphological evaluation of the embryos was conducted under a stereomicroscope using the morphologic scoring system by Brown and Fabro.The mitochondrial reactive oxygen species (ROS) level was detected by FACSCalibur flow cyto-metry using MitoSOXTM Red staining.Results: An obvious antagonistic effect was achieved through querce-tin at the dose of 1.0 μmol/L, which could result in an increase of visceral yolk sac (VYS) diameter, crown-rump length and head length, somite number, and the differentiation of visceral yolk sac vascular vessels.The scores of allantois, flexion, heart, hind brain, midbrain, forebrain, auditory system, visual system, olfactory system, branchialarch, maxillary process, forelimb bud and hindlimb bud also revealed a significant increase and the relative mitochondrial ROS level of embryonic cells was significantly decreased when compared with PM2.5 group.Although quercetin at the doses of 0.1 μmol/L, 0.5 μmol/L, 5.0 μmol/L also exhibited protective effects against PM2.5-induced embryonic developmental toxicity, the protective effect was weaker when compared with the dose of 1.0 μmol/L.Conclusion: Quercetin at proper dose may be of great benefit for the development of embryos exposed to PM2.5 in the uterus of the rats.Quercetin provides an effective strategy for the prevention of PM2.5-induced embryonic developmental toxicity.Clearance of mitochondrial ROS may be one of its mechanisms.