Development of polymorphic SSR markers in the razor clam (Sinonovacula constricta) and cross-species amplification.

Next-generation sequencing provides large-scale sequencing data with relative ease and at a reasonable cost, making it possible to identify a large amount of SSR markers in a timely and cost-effective manner. On the basis of the transcriptome database of Sinonovacula constricta obtained by Illumina/Solexa pyrosequencing, 60 polymorphic SSR markers were developed and characterized in 30 individuals. The number of alleles per polymorphic locus ranged from 2 to 7 with an average of 3.75 alleles. The observed and expected heterozygosities varied from 0.050 to 1.000 and from 0.050 to 0.836, respectively. Nineteen loci significantly deviated from Hardy-Weinberg equilibrium (P < 0.01) after Bonferroni's correction for multiple tests. In addition, interspecific transferability revealed that 20 polymorphic loci in Solen linearis were first characterized in this study. To the best of our knowledge, this is the highest number of SSRs in S. constricta and the first report of cross-species amplification. These novel polymorphic SSR markers will be particularly useful for conservation genetics, evolutionary studies, genetic trait mapping, and marker assisted selection in the species.

Effect of siRNA targeting HER2/neu on the proliferation and viability of prostate cancer PC-3M cells.

The aim of this study was to investigate the effect of a small interfering RNA (siRNA) targeting human epidermal growth factor receptor 2 (HER2/neu) on the proliferation and viability of prostate cancer PC-3M cells. Chemically synthesized siRNA targeting HER2/neu was transfected into PC-3M cells by using liposomes, and cells transfected with empty liposomes, a negative siRNA sequence, or nothing (untransfected) were used as controls. mRNA and protein levels of HER2/neu were detected using reverse transcription-polymerase chain reaction and western blot, respectively. The inhibitory action of HER2/neu siRNA on the in vitro growth of PC-3M cells was assessed by the cholecystokinin 8 assay and apoptosis was detected using flow cytometry. Cells transfected with HER2/neu siRNA showed decreased mRNA and protein levels of HER2/neu compared to control groups (P < 0.05). The survival rate of PC-3M cells decreased significantly after transfection with HER2/neu siRNA compared to that of untransfected cells (55.39 ± 1.60 and 81.42 ± 0.80%, respectively; P < 0.05). The apoptosis rate in cells transfected with HER2/neu siRNA was quite high (45.60 ± 0.70%) compared to that of blank control, empty liposome, and negative siRNA sequence groups (P < 0.05). In conclusion, siRNA targeting HER2/neu inhibits HER2/neu expression in PC-3M cells, resulting in an inhibition in proliferation and an induction of apoptosis.

Effect of Repeated Recruitment Manoeuvres on Patients with Severe Acute Respiratory Distress Syndrome.

OBJECTIVE: The study aimed to evaluate the influence of repeated recruitment manoeuvres (RRMs) on lung injury in patients with acute respiratory distress syndrome (ARDS). METHODS: Forty-one ventilated patients with severe ARDS were selected for this study. Recruitment manoeuvres (RMs) were conducted with continuous positive airway pressure (CPAP; 30 cm H2O for 40 seconds). Recruitment manoeuvres were repeated every two hours for all three groups. Changes in haemodynamics, pulmonary compliance, gas exchange and extravascular lung water index (EVLWI) were monitored before RM (pre-RM), 10 minutes after each RM, and four hours after RM3 (4 hours post-RRM). Pulmonary inflammatory factors (tumour necrosis factor-alpha [TNF-α] and interleukin [IL]-6 and -10) were also analysed. RESULTS: Compared with those in pre-RM, pulmonary compliance, oxygenation index (ratio of partial pressure of arterial oxygen to fraction of inspired oxygen [PaO2/FiO2]) and EVLWI remarkably improved in RM1, RM2, RM3 and 4 hours post-RRM (p < 0.05). The PaO2/FiO2 ratio increased significantly in RM1 and RM3 (p < 0.05). Extravascular lung water index decreased significantly in RM1 compared with that in RM3 and 4 hours post-RRM (p < 0.05). There was no significant difference in cytokines. CONCLUSION: Repeated recruitment manoeuvres during lung-protected ventilation can improve pulmonary compliance and oxygenation and significantly decrease extravascular lung water in ARDS patients. Lung injury was not worsened by RRMs in patients with severe ARDS.