Establishment and significance of pyrolysis model of human trophoblast HTR-8/SVneo induced by hydrogen peroxide / 医学研究生学报

ObjectiveA good invasion ability of extravilloustrophoblas (EVTs) is the prerequisite for successful placental colonization and effective remodeling of the uterine spiral artery. This article aims to simulate the pathophysiological process of oxidative stress inducing trophoblasts to pyroptosis in vitro, exploring the correlation between trophoblasts pyroptosis and the pathogenesis of preeclampsia.MethodsTwenty-five patients with preeclampsia were selected from the Department of Obstetrics and Gynecology, Zhongda Hospital affiliated to Southeast University from September 2017 to January 2019. Among them, early-onset preeclampsia (gestational weeks<34) was early-onset group (n=17), late-onset preeclampsia (gestational weeks≥34) was late-onset group (n=8), and full-term pregnant women with normal blood pressure (39＜gestational weeks＞42) were selected as normal group (n=10). Human trophoblasts were cultured with HTR-8/SVneo for 12 hours, and then treated with H2O2 (100, 150, 200, 250μmol/L) (2, 4, 6, 12 h), to induce human trophoblast HTR-8/SVneo pyrolysis model; the control group was normal cultured cells of 1640+10% fetal bovine serum + 1% antibiotics. Placental specimens from 7 patients with preeclampsia were randomly selected, including 3 cases in early onset group, 4 cases in late onset group and 1 case in normal group. The total proteins of cells and placenta were extracted respectively, and the expression of scorch death-related molecular proteins was detected. The mRNA levels of pyroptosis related molecules in cells was detected by RT-qPCR, and the morphological changes of cells were observed by inverted phase contrast microscope.ResultsThe Western blot results showed that the activation of the key molecular activation form of the cell pyrogenesis pathway, Cleaved caspase1, could be detected in the placenta. When H2O2 was 150 mol/L for 2h, the mRNA levels of NLRP3 and IL-1, the key molecules of the upstream activation signal, were significantly up-regulated (8.680±0.481, 14.136±0.244) compared with the control group (1.00±0.00) (P<0.000). At 4h, mRNA levels of key molecule GSDMD and downstream inflammatory factor IL-18 (1.639±0.354 and 1.794±0.043) in the pyrogenesis pathway were significantly higher than those in the control group (1.00±0.00), with statistically significant differences (P<0.05). By reverse validation of the mRNA levels of the molecules associated with pyroptosis, the optimal conditions of the model induced by H2O2 were 150 mol/L and 4h, and the typical changes, such as cell swelling, fragmentation and plasma membrane bubble formation, could be seen under the light microscope.ConclusionThe pyroptosis model of trophoblast cells was successfully established, and the physiological process of oxidative stress inducing trophoblasts to pyroptosis in vitro was successfully simulated, providing new ideas and directions for the diagnosis and treatment of preeclampsia and the development of new drugs.

Molecular Mechanism of CRBN in the Activity of Lenalidomid eagainst Myeloma--Review / 中国实验血液学杂志

Cereblon（CRBN) is a brain-associated protein with ionic protease activity, which interacts with DNA damage-binding protein-1 (DDB1), Cullin 4 (Cul4A or Cul4B), and regulator of Cullins 1 (RoC1) to form the functional E3 ubiquitin ligase complex(CRBN-CRL4) that performs proteolysis via the ubiquitin-proteasome pathway. And CRBN is a necessary target protein for the anti-myeloma effect of immunomodulators. The combination of lenalidomide and CRBN recruited a new substrate that binds to the CRBN-CRL4 complex, leading to increased ubiquitination and proteasome-dependent degradation, thus resulting in anti-myeloma activity. The substrates binding to this complex are IKZF1, IKZF3 proteins and GS, etc. The CRBN-dependent degradation of IKZF1 and IKZF3 after lenalidomide treatment is also the result of HO-mediated oxidative stress. In addition to ubiquitination, lenalidomide also mediates ubiquitin-independent pathways that prevent CRBN from binding to CD147-MCT1 in a competitive manner to regulate its antitumor activity. Lenalidomide can also play a role in multiple myeloma(MM) cells by modulating miRNA levels and CRBN binding to downstream protein AGO2 expression. Thus, there are many molecular mechanisms of lenalidomide anti-myeloma activity. This review summarizes the molecular mechanisms of CRBN in lenalidomide against myeloma activity in terms of ubiquitin-dependent and ubiquitin-independent pathways.

Participatory Teaching Class for MBBS Students / 昆明医科大学学报

Objective To determine the effect of participatory teaching method in the Clinic teaching of internal medicine for MBBS students. Methods The 60 MBBS students were divided into group A and B, 30 students in each group. Participatory teaching was done in group A, and traditional teaching was carried out in group B,and the effects of the two types of teaching methods were compared. Results In the clinic teaching of internal medicine, the effects in improving students' clinical thinking and practical ability, creativeness, initiative and efficiency were better in the participatory teaching method than in traditional teaching method (P＜0.05) . Conclusion The effects of participatory teaching in the clinic teaching of internal medicine for MBBS students was better than the traditional teaching.

The Value of Using sPESI Score Combined with hs-TnT in the Evaluation of Short-term Prognosis in Hemodynamically Stable Pulmonary Embolism / 昆明医科大学学报

Objective To explore the value of using sPESI score and hs-TnT in the evaluation of short-term prognosis in hemodynamically stable pulmonary embolism. Methods We collected 99 patients with hemodynamically stable PE from our department. According to the sPESI score and hs-TnT, patients were divided into high risk group (sPESI score≥1) and low risk group (sPESI score=0), positive group (hs-TnT≥0.014 ng/mL) and negative group (hs-TnT＜0.014 ng/mL) . Then all patients were treated and followed up for 30 days. The sensitivity, specificity, positive predictive value and negative predictive value of adverse events of prognosis were calculated, and ROC curve was drawn to analyze the values in different grouping methods for the prognostic evaluation.Results Thirteen adverse events occurred in all patients. The single test showed that sensitivity, specificity, positive predictive value, and negative predictive value of hs-TnT for predicting adverse events were respectively 84.6%,55.2%,22%,and 96%. When sPESI was used alone, sensitivity, specificity, positive predictive value and negative predictive value were 92.3%, 48.8%, 21.1% , and 97.7% . Results of combined testing showed 100% sensitivity, 29% specificity, 17.6% positive predictive value, and 100% negative predictive value. The ROC curve area of the sPESI, hs-TnT, sPESI and hs-TnT are 0.832 (95% CI, 0.705-0.958), 0.825 (95%CI, 0.694-0.957),0.872 (95%CI, 0.773-0.971) . Conclusions PESI and hs-TnT have clinical value in evaluating the short-term prognosis of hemodynamically stable pulmonary embolism. sPESI combined with hs-TnT has higher significance, especially in patients with low-risk PE.

Antioxidant effect of salvianolic acid B on hippocampal CA1 neurons in mice with cerebral ischemia and reperfusion injury / 中国结合医学杂志

<p><b>OBJETIVE</b>To investigate the neuroprotective effects and underlying mechanisms of salvianolic acid B (Sal B) extracted from Salvia miltiorrhiza on hippocampal CA1 neurons in mice with cerebral ischemia reperfusion injury.</p><p><b>METHODS</b>Forty male National Institute of Health (NIH) mice were randomly divided into 4 groups with 10 animals each, including the sham group, the model group, the SalB group (SalB 22.5 mg/kg) and the nimodipine (Nim) group (Nim 1 mg/kg). A mouse model of cerebral ischemia and reperfusion injury was established by bilateral carotid artery occlusion for 30 min followed by 24-h reperfusion. The malondialdehyde (MDA) content, the nitric oxide synthase (NOS) activity, the superoxide dismutase (SOD) activity and total antioxidant capability (T-AOC) of the pallium were determined by biochemistry methods. The morphologic changes and Bcl-2 and Bax protein expression in hippocampal CA1 neurons were observed by using hematoxylineosin staining and immunohistochemistry staining, respectively.</p><p><b>RESULTS</b>In the SalB group, the MDA content and the NOS activity of the pallium in cerebral ischemia-reperfusion mice significantly decreased and the SOD activity and the T-AOC significantly increased, as compared with the model group (P<0.05 or P<0.01). The SalB treatment also rescued neuronal loss (P<0.01) in the hippocampal CA1 region, strongly promoted Bcl-2 protein expression (P<0.01) and inhibited Bax protein expression (P<0.05).</p><p><b>CONCLUSIONS</b>SalB increases the level of antioxidant substances and decreases free radicals production. Moreover, it also improves Bcl-2 expression and reduces Bax expression. SalB may exert the neuroprotective effect through mitochondria-dependent pathway on hippocampal CA1 neurons in mice with cerebral ischemia and reperfusion injury and suggested that SalB represents a promising candidate for the prevention and treatment of ischemic cerebrovascular disease.</p>

Effects of Haikun Shenxi on expression of platelet-derived growth factor-B and mRNA in renal tissue of rats with adriamycin nephropathy / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effects of Haikun Shenxi on the expression of platelet-derived growth factor-BB (PDGF-BB) and mRNA in renal tissue of rats with adriamycin nephropathy.</p><p><b>METHOD</b>Rat model was established by unilateral nephrectomy and injecting adriamycin intraperitoneally. The adriamycin-induced nephrotic rats were randomly divided into 6 groups: normal group, sham operation group, model group, lotensin treatment group, Haikun Shenxi low and high dose treatment groups (0.77, 0.08 mg x kg(-1). Ten weeks later, the 24 hour urine protein and blood biochemistry examinations and renal pathologic changes were observed, and the expression of PDGF-BB and mRNA was measured using immunohistochemical method.</p><p><b>RESULT</b>Compared with model group, proteinuria and the levels of serum creatinine (Scr) , urea nitrogen (BUN) were decreased obviously in both Haikun Shenxi low and high dose groups. The expression of PDGF-BB and mRNA was mostly presented in cytoplasm of renal tubular epithelial cells and mesangial area, and it could be reduced significantly after treatment (P < 0. 05).</p><p><b>CONCLUSION</b>The level of PDGF-BB and mRNA is high in renal tissue of adriamycin-induced nephrotic rats. This progress could be effectively inhibited by Haikun Shenxi and the mechanism may be that it can control the excessive expression of PDGF-BB and mRNA.</p>

Rat alveolar type II injured by bleomycin / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore dysfunction mechanism of rat alveolar type II (AT-II) injured by bleomycin (BLM).</p><p><b>METHODS</b>SD rats were injected with a single intratracheal dose of bleomycin or control saline. On day 7, 14, and 28 following intratracheal bleomycin or saline instillation, animals were killed under overdose of 1.5% sodium pentobarbital (0.25 ml/100 g, i.p.) and bronchoalveolar lavage fluid (BALF) from the lung was tested for the activity of pulmonary surfactant (PS) by the Whihelmy Film Balance. Several concentrations of bleomycin stimulated the culture of rat AT-II cells, and surfactant protein (SP) A, B, and aquaporin-1 (AQP) mRNA were analyzed by fluorescent quantitative polymerase chain reaction (FQ-PCR).</p><p><b>RESULTS</b>The activity of PS and hypoxemia significantly decreased on day 7 and improved on day 14 and completely recovered to normal status on day 28. SP-A, B, and AQP-1 mRNA expression in BLM-stimulated group were significantly lower than those in the control group (P<0.001).</p><p><b>CONCLUSION</b>BLM-injured AT-II cells decrease the levels of SP-A, B, and AQP-1 mRNA and cause PS dysfunction, resulting in hypoxemia and pneumonedema.</p>

Influence of compound biejia ruangan prescription on extracelluar matrix in bleomycin induced pulmonary fibrosis rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the influence of compound Biejia Ruangan Prescription (CBRP) on extracelluar matrix in bleomycin induced pulmonary fibrosis rats.</p><p><b>METHOD</b>54 male Sprague-Dawley rats were randomly divided into 6 groups (9 rats in each group). Rats in the model control group, positive medicine group, and high, moderate and low CBRP groups were injected with bleomycin A5 by trachea, and rats in sham-model control group with same volume normal saline. 29 days after the injection, CBRP solution of different dosages (1.4 g x kg(-1), 0.7 g x kg(-1), 0.35 g x kg(-1)) was respectively given to rats in the high, moderate and low CBRP group by gavage, while equal volume of normal saline was given to those in the sham-model control group and model control group, and an equal volume of prednisone (0.56 mg x kg(-1)) was given to those in positive medicine control group. On the 80th day, the levels of III-collagen, IV-collagen, laminin and hyaluronic acid in the serum were determined, the determination of hydroxyproline in lung homogenates was analyzed, and the right lung was incised to make pathological sections which were stained with Hematoxylin-Eosin (HE) and Masson staining for pathological diagnosis.</p><p><b>RESULT</b>CBRP could decrease the levels of III-collagen, IV-collagen, laminin and hyaluronic acid in the serum.</p><p><b>CONCLUSION</b>CBRP may play its therapeutic role by leveling down the content of extracellular matrix in rats with pulmonary fibrosis induced by Bleomycin A5.</p>

Study on the interface of human hepatocyte L-02 polypropylene:simple culture method of human hepatocyte with spheroidal aggregate culture / 中华外科杂志

<p><b>OBJECTIVE</b>To found new interface of human hepatocyte/poly propylene with good cytocompatibility for made polypropylene hollow fibers bioreactor of bioartificial liver in future.</p><p><b>METHODS</b>Using the macromolecular hydroperoxide groups on the polypropylene membrane surface as initiators, acrylamides were polymerized on the polypropylene membranes, under induction by both UV irradiation and Fe2+ reduction. Growth characteristics of human hepatocyte L-02 were detected when it was cultured on polystyrene, polypropylene and modified polypropylene membrane surface.</p><p><b>RESULTS</b>Water contact angle measurement of the polypropylene and the modified polypropylene membranes decreased from (72 +/- 5) degrees to (30 +/- 4) degrees , which indicated that the hydrophilicity of the membrane was improved obviously after the grafting modification. Human hepatocyte L-02 could not adhere and spread on modified polypropylene membrane surface, and grown in spheroidal aggregate with higher density and higher proliferation ratio measured by MTT method.</p><p><b>CONCLUSIONS</b>Acrylamide polymerized on the polypropylene membranes is a good method which not only improved human hepatocytes cytocompatibility but also found a new simple culture method with spheroidal aggregate culture of human hepatocyte.</p>