A new species of the genus Megophrys (Amphibia: Anura: Megophryidae) from Hunan, China.

We describe a new species of Megophryidae frog, Megophrys shunhuangensis sp. nov., from Hunan Province, Southern China. It can be distinguished from other known species in the subgenus Panophrys by morphological characters, bioacoustic data and a molecular divergence in the mitochondrial 16S rRNA gene similar to that found among other species of Panophrys. M. shunhuangensis sp. nov. is characterized by a relatively small body size, with adult females measuring 37.6 mm and adult males measuring 30.3-33.6 mm in snout to vent length; maxillary teeth present, vomerine teeth absent; tongue smooth, not notched behind; hindlimb slender, heels overlapping, tibio-tarsal articulation reaches forward between the nasal and tip of snout. Molecular phylogenetic analyses also show that M. sp. 6 from (Mao'er Shan, China) from Chen et al. (2017) and M. sp. 24 (Anjiangping and Mao'er Shan, China) from Liu et al. (2018) may be the same species as M. shunhuangensis sp. nov., we consequently speculate that the new species also distributed in Anjiangping and Mao'er Shan, China. At present, the genus Megophrys contains 85 species of which 48 species are distributed in China, and 30 belong to the subgenus Panophrys.

Functional analyses of Populus euphratica brassinosteroid biosynthesis enzyme genes DWF4 (PeDWF4) and CPD (PeCPD) in the regulation of growth and development of Arabidopsis thaliana.

DWF4 and CPD are key brassinosteroids (BRs) biosynthesis enzyme genes. To explore the function of Populus euphratica DWF4 (PeDWF4) and CPD (PeCPD), Arabidopsis thaliana transgenic lines (TLs) expressing PeDWF4, PeCPD or PeDWF4 plus PeCPD, namely PeDWF4-TL, PeCPD-TL and PeCP/DW-TL, were characterized. Compared with wild type (WT), the changes of both PeDWF4-TL and PeCPD-TL in plant heights, silique and hypocotyls lengths and seed yields were similar, but in bolting time and stem diameters, they were opposite. PeCP/DW-TL was more in plant heights and the lengths of primary root, silique, and fruit stalk, but less in silique numbers and seed yields than either PeDWF4-TL or PeCPD-TL. PeDWF4 and PeCPD specially expressed in PeDWF4-TL or PeCPDTL, and the transcription level of PeDWF4 was higher than that of PeCPD. In PeCP/DW-TL, their expressions were all relatively reduced. Additionally, the expression of PeDWF4 and PeCPD differentially made the expression levels of AtDWF4, AtCPD, AtBR6OX2, AtFLC, AtTCP1 and AtGA5 change in the TLs. The total BRs contents were PeDWF4-TL greater than PeCP/DW-TL greater than WT greater than PeCPD-TL. These results imply that PeDWF4 is functionally not exactly the same as PeCPD and there may be a synergistic and antagonistic effects in physiology between both of them in the regulation of plant growth and development.

Circulating microRNAs indicate cardioprotection by sevoflurane inhalation in patients undergoing off-pump coronary artery bypass surgery.

In patients undergoing off-pump coronary artery bypass surgery (OPCAB), it is important to attenuate myocardium injury during the surgery. The present study aimed to observe the cardioprotection induced by sevoflurane induction and maintenance compared with propofol intravenous anesthesia, and to detect its potential protection against acute myocardial injury with sensitive biomarkers. In total, 36 patients undergoing OPCAB were randomly assigned into two groups, receiving sevoflurane (n=18) or propofol (n=18) as the induction and maintenance anesthetic agent. The depth of anesthesia in the two groups was kept at a bispectral index value of 40-50. Physiological and hemodynamic parameters were recorded during the surgery. Cardiac troponin-I (cTnI), creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH) and two microRNAs (miR-499 and miR-208b) were also measured during and subsequent to surgery. Nno statistically significant differences were observed in the physiological and hemodynamic parameters between the two groups prior to surgery. Following surgery, the cardiac output and stroke volume improved significantly in the sevoflurane group (P<0.05). In addition, patients in the sevoflurane group had lower miR-499 (P<0.05) and miR-208b (P<0.01) levels at 12 h after surgery when compared with the propofol group. However, no significant differences in cTnI, CK-MB and LDH levels were observed following surgery between the two groups. In conclusion, volatile induction and maintenance with sevoflurane resulted in some extent of cardiac function improvement in patients undergoing OPCAB. Cardioprotection by sevoflurane is suggested by reduced cardiac injury compared with propofol, and indicated by the sensitive biomarkers, circulating miR-499 and miR-208b.

Premature rupture of the fetal membrane combined with subclinical chorioamnionitis negatively affects pregnancy outcomes by a mechanism associated with reduced levels of matrix metalloproteinase-2.

The present study aimed to investigate whether premature rupture of the fetal membrane, combined with subclinical chorioamnionitis, affects pregnancy outcomes. In addition, the association between premature rupture of the fetal membrane (PROM) and the levels of matrix metalloproteinase-2 (MMP-2), an inactive proenzyme that can be activated by other factors or signals in humans, was examined. In total, 80 pregnant patients with PROM were classified into the experimental and control groups, according to their final placental pathological diagnosis results. The 40 patients in the experimental group suffered from subclinical chorioamnionitis, while the 40 patients in the control group exhibited no lesions of the placenta or fetal membrane. Tissue samples were collected and the total protein and mRNA were extracted for western blot and quantitative polymerase chain reaction analyses. ELISA was performed in order to detect the levels of MMP-2 in the serum of the two groups of patients. The rates of cesarean section, puerperal infection, postpartum hemorrhage, preterm incidence, placenta accreta, residual placental blood and stillbirth were all significantly higher in the experimental group compared with the control group. In addition, the mRNA and protein expression levels of MMP-2 were reduced in the experimental group compared with the control group. ELISA results indicated that the serum MMP-2 concentrations were also reduced in the patients with PROM. Therefore, the present study demonstrated that the PROM, combined with subclinical chorioamnionitis, significantly affected pregnancy outcomes and was associated with reduced levels of MMP-2.

Effects of paroxetine on spatial memory function and protein kinase C expression in a rat model of depression.

The aim of the present study was to investigate the effects of paroxetine on the spatial memory and expression level of protein kinase C (PKC) in a rat model of depression. Rat models of depression were established by chronic unpredictable mild stress. The spatial learning and memory function of the rats were assessed by the Morris water maze test. The expression levels of PKC in the hippocampus were detected by western blotting. The results showed that, compared with the control group, the escape latency was prolonged and the percentage of time in the target quadrant and the number of times the rats crossed the platform were reduced in the model group; however, the impaired spatial learning and memory function in these rat models could be restored by paroxetine, almost to a level comparable with that in the normal control animals. In addition, the expression of PKC in the model group was significantly decreased compared with that in the control group, and the expression could also be elevated by paroxetine treatment. These results suggest an association between PKC levels and the pathogenesis of depression. The application of paroxetine can improve the spatial memory and reverse the changes in PKC levels in the hippocampus in the rat model of depression. The present findings have enhanced the understanding of the pathogenesis of depression, and provide experimental evidence for the treatment of depression with paroxetine.

Endocrine MPA enhances the effects of TAC chemotherapy on improvement of prognosis and increase in long-term survival rates for patients with endometrial cancer.

The aim of the present study was to investigate the effect of taxol, adriamycin and carboplatin (TAC) chemotherapy combined with endocrine medroxyprogesterone acetate (MPA) therapy for the treatment of patients with endometrial cancer. A retrospective analysis of 124 patients with endometrial cancer was performed by dividing the cohort into an experimental and control group. The 64 patients in the experimental group received TAC and MPA chemotherapy, whereas the 60 patients in the control group were treated with TAC chemotherapy only. Tissue samples scraped from the uterus were used to extract the total proteins and RNAs for the western blot and reverse transcription-quantitative polymerase chain reaction analyses, respectively. All the patients were followed up for 20-45 months, during which time prognostic data, and one- to three-year survival rates were recorded and compared. The rate of recurrence or metastasis was significantly lower in the experimental group compared with that in the control group (P<0.05) and the three-year survival rate of the experimental group was significantly higher than that of the control group (P<0.05). Furthermore, the mean metastasis-associated 1 (MTA1) protein and RNA expression levels were significantly lower in the experimental group compared with the control group (P<0.05), exhibiting ~30 and ~15% of the levels in the control group, respectively. Therefore, a treatment strategy of TAC chemotherapy combined with endocrine MPA therapy appears to effectively improve the prognosis and increase the long-term survival rates of patients with endometrial cancer. Such an enhancing effect may be mediated by the transcriptional downregulation of MTA1 expression.

Role of autophagy in the ω-3 long chain polyunsaturated fatty acid-induced death of lung cancer A549 cells.

The present study identified that ω-3 long chain polyunsaturated fatty acids (ω-3 PUFAs), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) demonstrate anti-proliferative effects in lung cancer A549 cells. MTS and cytotoxicity assays were conducted to confirm that ω-3 PUFAs induced cell death. Autophagy-associated gene and signaling pathways were also detected. Microtubule-associated protein light chain 3 (LC3) expression was found to be increased subsequent to treatment with DHA and EPA, and the expression of LC3-II was particularly increased. mRFP-GFP-LC3 fluorescence staining and p62 expression levels were used to detect autophagic flux. The present results indicate that DHA and EPA block autophagic flux, suggesting autophagosome accumulation. Subsequent to treatment with DHA and EPA, which interfered with autophagosomes, the expression of Beclin 1 was significantly decreased, while the expression of phosphorylated Akt and phosphorylated mammalian target of rapamycin was significantly increased. Therefore, DHA and EPA exert anti-proliferative effects by inhibiting autophagy in A549 cells, which highlights the potential of DHA and EPA for use in the prevention or treatment of lung cancer.

Levels of plasma Epstein-Barr virus DNA prior and subsequent to treatment predicts the prognosis of nasopharyngeal carcinoma.

The level of Epstein-Barr virus DNA (EBV-DNA) in the plasma prior and subsequent to treatment is a reliable biomarker for the screening, diagnosis, monitoring and prognosis of nasopharyngeal carcinoma (NPC). The present retrospective study aimed to determine whether pre- and post-treatment levels of plasma EBV-DNA were predictive of survival in a large sample of patients with NPC. The level of plasma EBV-DNA in 637 NPC patients prior and subsequent to treatment was determined by quantitative polymerase chain reaction. The value of pre- and post-treatment plasma EBV-DNA in predicting the survival of NPC patients was then analyzed. The results revealed that pre-treatment plasma EBV-DNA loads were significantly higher in patients with NPC than those in healthy controls (P<0.001). The percentage of patients with positive plasma EBV-DNA was markedly higher prior to treatment (70.64%; median, 1150 copies/ml; range, 0-9.75×106 copies/ml) than following treatment (25.99%; median, 0 copies/ml; range, 0-3.83×106 copies/ml) (P<0.001). Patients with a high plasma EBV-DNA load presented with a higher clinical tumor classification, lymph node status, metastatic status and overall cancer stage. The risk of NPC relapse and mortality was higher in patients with pre-treatment plasma EBV-DNA levels of ≥1,500 copies/ml than that in patients with <1,500 copies/ml. Furthermore, the risk of relapse and mortality was higher in patients with positive post-treatment plasma EBV-DNA than in patients with negative post-treatment plasma EBV-DNA. Detectable post-treatment plasma EBV-DNA was the most significant prognostic factor to affect relapse-free survival, whilst metastasis was the prognostic factor with the greatest effect on overall survival. These data indicated that pre- and post-treatment levels of plasma EBV-DNA were able to predict the prognosis of NPC. This finding may provide novel references for research and clinical practice.

Effects of Jin-Ying-Tang on Staphylococcus aureus-induced mastitis in rabbit.

The present study was performed to investigate the effects of Jin-Ying-Tang (JYT), a Chinese herbal formula containing Lonicera japonica, Herba taraxaci, Fructus trichosanthis, Fructus forsythia, Radix et rhizoma rhei, Astragalus membranaceus, Angelica sinensis, on rabbit mastitis induced by Staphylococcus aureus. Suckling rabbits were challenged with 1.5 × 10(7) colony forming unit (CFU) of S. aureus at the base of the third pair teats, and they were treated and pretreated with JYT to detect the formula effects. The results showed that JYT could reduce the occurrence of Staphylococcal mastitis in rabbit model. To further investigate the action mechanism of JYT, we examined the leukocyte counts and inflammatory mediator levels such as TNF-α and IL-6 in blood and infected tissue. From histological study and blood analysis, we found that JYT could suppress leukocyte infiltration in infected mammary gland tissue and significantly inhibit the total leukocyte counts and lymphocytes (LYM), monocytes (MON) and granulocytes (GRA) fractions of leukocyte counts in blood. Enzyme-linked immunosorbent assay (ELISA) results showed JYT significantly decreased the TNF-α and IL-6 concentrations in serum and mammary gland. The analysis of these data suggested that JYT effectively inhibited inflammatory responses to reduce the occurrence of mastitis in rabbit model.

Anticoagulation in patients with heparin-induced thrombocytopenia type II.

Heparin-induced thrombocytopenia (HIT) together with simultaneously occurring thromboembolism is a serious complication of heparin treatment. At present an immunologic cause of this side effect of heparin is equivocally accepted. However, further anticoagulation of these patients is still debated. The present article summarizes the treatment of 20 patients with such complications. Two of these patients did not develop thrombocytopenia but presented cutaneous allergy or necrosis. All patients were treated either with intravenous heparinoid (Orgaran) or with low-molecular-weight heparin without/with simultaneous intravenous high-dose immunoglobulins or with intravenous r-hirudin. Based on these experiences the treatment of choice depends at present on the availability of the anticoagulants and on the local experience with the different anticoagulants. In the future r-hirudins and other nonheparin thrombin inhibitors may become the drugs of choice in this indication. Surgical intervention has to be considered additionally.