The inflammatory response is involved in the pathogenesis of the most common types of heart disease.Sanguinarine (SAN) has various pharmacological properties such as anti-inflammatory,antioxidant,antibacterial,antitumor,and immune-enhancing properties.However,few studies have investigated the effects of SAN on lipopolysaccharide (LPS)-induced inflammatory and apoptotic responses in H9c2 cardiomyocytes.Therefore,in this study,H9c2 cells were co-treated with SAN and LPS,and the mRNA levels of pro-inflammation markers and the apoptosis rate were measured to clarify the effect of SAN on cardiac inflammation.The underlying mechanism was further investigated by detecting the activation of Toll-like receptor (TLR)4/nuclear factor-κB (NF-κB) signaling pathways.As a result,increased mRNA expression of interleukin (IL)-1 β,IL-6,and TNFα induced by LPS was attenuated after SAN treatment;LPS-induced apoptosis of H9c2 cardiomyocytes and cleaved-caspase 8,9,3 were all significantly reduced by SAN.Further experiments showed that the beneficial effect of SAN on blocking the inflammation and apoptosis of H9c2 cardiomyocytes induced by LPS was associated with suppression of the TLR4/NF-κB signaling pathway.It was suggested that SAN suppressed the LPS-induced inflammation and apoptosis of H9c2 cardiomyocytes,which may be mediated by inhibition of the TLR4/NF-κB signaling pathway.Thus,SAN may be a feasible therapy to treat sepsis patients with cardiac dysfunction.

Sequential methodology based on the application of three types of experimental designs was used to optimize the fermentation conditions for elastase production from mutant strain ZJUEL31410 of Bacillus licheniformis in shaking flask cultures. The optimal cultivation conditions stimulating the maximal elastase production consist of 220 r/min shaking speed, 25 h fermentation time, 5% (v/v) inoculums volume, 25 ml medium volume in 250 ml Erlenmeyer flask and 18 h seed age. Under the optimized conditions, the predicted maximal elastase activity was 495 U/ml. The application of response surface methodology resulted in a significant enhancement in elastase production. The effects of other factors such as elastin and the growth factor (corn steep flour) on elastase production and cell growth were also investigated in the current study. The elastin had no significant effect on enzyme-improved production. It is still not clear whether the elastin plays a role as a nitrogen source or not. Corn steep flour was verified to be the best and required factor for elastase production and cell growth by Bacillus licheniformis ZJUEL31410.
Bacillus ; enzymology ; growth & development ; Fermentation ; Pancreatic Elastase ; biosynthesis ; Research Design

ObjectiveTo investigate the value of 18F-FDG SPECT myocardial imaging in evaluating haemodynamic change,treatment outcome and prognosis for idiopathic pulmonary arterial hypertension (IPAH).MethodsAll 24 patients with IPAH underwent 18 F-FDG SPECT myocardial imaging.Right ventricle/left ventricle (RV/LV)-FDG uptake was calculated by ROI method drawing over the central areas of left and right ventricular free walls.All patients underwent right heart catheterization within 3 days after imaging studies.Mean pulmonary artery pressure (mPAP) and pulmonary vascular resistance (PVR) were recorded.After six month pharmaceutical treatment,15 IPAH patients were re-examined with 18F-FDG SPECT myocardial imaging followed by repeated right heart catheterization within 3 days.Plasma N-terminal pro-brain naturetic peptide (NT-proBNP) and endothelin-1 ( ET-1 ) were measured in 17 patients using electrochemiluminescent immunoassay and enzyme immunoassay respectively.All patients were followed up for 12 months at least.Correlations between RV/LV-FDG uptake and mPAP and PVR were determined by simple linear regression analysis.Change of RV/LV-FDG before and after treatment was calculated using Student's t-test.Survival in groups with RV/LV FDG uptake ≥ 1.15 and RV/LV-FDG uptake ＜ 1.15 were compared using Log-rank test.ResultsSignificant correlations were found between RV/LV-FDG uptake and mPAP (r =0.562,P ＜ 0.01 ),and between RV/LV-FDG uptake and PVR ( r =0.574,P ＜ 0.01 ).There were no significant correlation between RV/LV-FDG uptake and NT-proBNP( r =0.18 1,P ＞ 0.05 ),but a significant correlation between RV/LV-FDG and ET-1 was observed (r =0.669,P ＜ 0.01 ).The RV/LV-FDG uptake in patients with positive treatment outcome ( n =6) decreased from 1.38 ± 0.52 to 0.92 ±0.26 (t =4.018,P ＜ 0.05) after 6 months treatment.In contrast,no significant change of RV/LV-FDG uptake was seen in those patients (n =9) with negative treatment outcome ( t =1.861,P ＞ 0.05 ).The mean follow-up time was (21 ±8) months.Mean survival time for the patients with RV/LV- FDG uptake ≥ 1.15was 28 months (95％ confidence interval:24-32 months),which was significantly lower than 34 months survival (95％ confidence interval:33-35 months) for the patients with RV/LV-FDG ＜ 1.15 (x2 =3.956,P ＜0.05 ).Conclusions Detection of right ventricle myocardial glucose metabolism level with 18F-FDG SPECT may be a practical method for evaluating haemodynamic change,treatment outcome and prognosis of IPAH.

The breeding of domestic rabbits (Oryctolagus cuniculus) for human consumption has a long tradition in China. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animal health. Thus, a total of 1,132 domestic rabbit sera from 4 regions in China were collected for serological screening for Encephalitozoon cuniculi and for Toxoplasma gondii by ELISA and modified agglutination test (MAT), respectively. Antibodies to E. cuniculi were detected in 248/1,132 (21.9%) sera tested while antibodies against T. gondii revealed a seroprevalence of 51/1,132 (4.5%). We believe that the present results are of epidemiological implications and public health importance due to the acknowledged susceptibility of humans to E. cuniculi and T. gondii infections. Therefore, routine screening tests of domestic rabbits are proposed considering the zoonotic potential of these parasites.
Animals ; Animals, Domestic/blood/microbiology/parasitology ; Antibodies, Fungal/*blood ; Antibodies, Protozoan/*blood ; China/epidemiology ; Encephalitozoon cuniculi/*immunology/isolation & purification ; Encephalitozoonosis/blood/microbiology/*veterinary ; Female ; Male ; Rabbits/blood/microbiology/parasitology ; Seroepidemiologic Studies ; Toxoplasma/*immunology/isolation & purification ; Toxoplasmosis, Animal/*blood/parasitology

Amino Acid Sequence ; Base Sequence ; Genes, Bacterial ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Staphylococcus epidermidis ; enzymology ; genetics ; beta-Lactamases ; chemistry ; genetics

This study was purposed to explore the changes of possible angiogenetic factors other than VEGF after inhibition of NHE1 and their related mechanisms. The K562 cells were treated by NHE1 specific inhibitor cariporide, the angiogenesis factors after inhibition of NHE1 were screened by using protein chip, the IL-8 expression level after cariporide treatment was detected by real-time quantitative PCR; the K562 cells with stable interference of NHE1 were constructed, the IL-8 expression level after interference of NHE1 was detected by real-time quantitative PCR; the p38 phosphorylation level in K562 cells treated with cariporide was detected by Western blot. After treatment of K562 cells with p38 inhibitor SB203580, the IL-8 expression level was decreased by real-time quantitative PCR. The results of protein chip showed that IL-8 expression decreased after cariporide treatment. Real-time quantitative PCR confirmed this inhibitory effect. The p38 phosphorylation level increased after cariporide treatment. The down-regulation of IL-8 expression induced by cariporide treatment was partially restored after K562 cells were treated with p38 inhibitor SB203580. It is concluded that the inhibition of NHE1 can inhibit IL-8 expression through up-regulation of p38 phosphorylation.
Cation Transport Proteins ; antagonists & inhibitors ; Down-Regulation ; Guanidines ; pharmacology ; Humans ; Imidazoles ; pharmacology ; Interleukin-8 ; metabolism ; K562 Cells ; Phosphorylation ; drug effects ; Pyridines ; pharmacology ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers ; antagonists & inhibitors ; Sulfones ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the association of the clinical characteristics and the outcome of in vitro fertilization and embryo transfer (IVF-ET) in infertile women with polycystic ovarian syndrome (PCOS) of different subtypes.

METHODSA total of 189 infertile women with PCOS undergoing IVF-ET were enrolled in this study. According to Rotterdam PCOS diagnosis criteria, the patients were classified into 3 PCOS subtypes, namely type I with PCO ultrasonography and oligo-ovulation/anovulation and hyperandrogenism (54 women, for whom 58 fresh IVF-ET cycles were performed); type II with PCO ultrasonography and oligo-ovulation/anovulation (117 women with 126 cycles); type III with PCO ultrasonography and hyperandrogenism (18 women with 18 cycles). The number of retrieved oocytes, fertilization rate, implantation rate, clinical pregnancy rate, spontaneous abortion rates and incidence of ovarian hyperstimulation syndrome (OHSS) were compared between the 3 groups.

RESULTSExcept for the baseline serum T concentration in the early phase of menstrual cycle, which was significantly higher in groups I and III than in group II, no significant difference was found in the clinical characteristics between the 3 groups (P>0.05). Group I had the highest initial Gn dose, and the oocyte retrieval rates were significantly lower in groups I and III (P<0.05). The patients in group I had lower implantation rate and the clinical, on-going and cumulative pregnancy rates than groups II and III, but the differences were not statistically significant; the embryo early loss rate and spontaneous abortion rate appeared to be higher in groups I and III (P>0.05). Significantly elevated incidence of OHSS were noted in groups I and III (P<0.05).

CONCLUSIONThe women with different PCOS subtypes according to the Rotterdam criteria all have similar IVF-ET outcomes, and the increased embryo loss rate and spontaneous abortion rate in groups I and III might be associated with excessive androgen that disturbs oocyte and embryo development.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; etiology ; therapy ; Polycystic Ovary Syndrome ; classification ; complications ; Pregnancy ; Pregnancy Rate

OBJECTIVETo explore the quality of the embryos, clinical outcomes and birth defects resulting from intracytoplasmic sperm injection (ICSI) treatment using sperms of different origins and parameters.

METHODSA total of 980 ICSI-ET (embryo-transfer) cycles were divided into 4 groups, namely normal sperm or mild oligozoospermia group (group A), severe oligozoospermia group (group B), epididymal aspirates group (group C) and testicular biopsies group (group D). The cleavage rate, embryo quality, fertilization rate, clinical pregnancy rate, and rates of birth defects were compared between the groups.

RESULTSThe fertilization rate, cleavage rate and good-quality embryo rate were not significantly different among the 4 groups (P>0.05), and the embryo implantation rate and clinical pregnancy rate were significantly higher in group C than in groups A and B (P<0.05). Groups A, B and C showed no significant differences in the rates of birth defects (P>0.05), and no birth defects occurred in group D.

CONCLUSIONSSperms of different parameters and origins used in ICSI treatment can achieve similar fertilization rate, good-quality embryo rate and delivery rate. The embryo implantation rate and clinical pregnancy rate of epididymal sperm group are higher than those of ejaculated groups, possibly due to the younger age of the patients and a greater number of oocytes retrieved in group C than in groups A and B.

Congenital Abnormalities ; etiology ; Female ; Fertilization ; Humans ; Male ; Sperm Injections, Intracytoplasmic ; adverse effects ; Spermatozoa ; Treatment Outcome

Objective To establish the mouse embryonic fibroblast (MEF) cells lines with stable expression of MiR-100-3p inhibitor,which lays a foundation for its function research of the role of miR-100 in the interaction of environmental pollutants and Src homology 2 domain tyrosine phosphatases (SHP-2) gene mutation.Methods Designed and synthesized a particular DNA fragment as hsa-MiR-100-3p inhibitor,which was then cloned into the hU6-MCS-Ubiquitin-ECFP -IRES-puromycin vector.This vector was then co-transfected with lentiviral packaging vectors pHelper1.0 and pHelper2.0 into 293T to produce lentiviral particles.Finally,MEF cells were infected with the above-mentioned lentiviral particles and the stable cell lines expressing hsa-miR-100-3p inhibitor were selected by puromycin.The expression of green fluorescent protein (GFP) and the expression of miR-100 were detected which indicated its inhibition effect of lentiviral vector in MEF cells.Results The construction of the recombinant plasmid was confirmed by sequencing.The subsequent infection confirmed that the packaging of lentiviruses was successfully.MEF cells successfully infected with hsa-miR-100-3p inhibitor.Flow cytometry indicates the expression of green fluorescent protein (significantly inhibition the expression of miR-100) has reached 97.4％ and 95.1％,as well as proved by RT-PCR.Conclusion We successfully constructed the has-miR-100-3p lentiviral silencing vector,and established the stable inhibition cell lines which lay a foundation for its function research of miR-100.

OBJECTIVE To explore the clinical effects of Shuangxin Decoction in treating coronary heart disease accompa-nied with psychological disorder.METHODS 80 cases with coronary heart disease manifested as chest pain,chest distress plus psychological disorders like depression and anxiety were randomly divided into treatment group and control group.Patients in both groups were treated with routine western medicine plus psychological counseling.Based on which,patients in the treat-ment group were further given Chinese medicine Shuangxin Decoction,while those in the control group were added deanxit, with the course lasting for 6 weeks.Chest pain(distress) integral score scale,traditional Chinese medicine(TCM) syndrome score scale and comprehensive hospital anxiety and depression scale were evaluated to explore the therapeutic effects of chest pain,psychological disorder and clinical TCM syndromes before and after treatment of both groups,together with the changes of electrocardiogram test results and cardiac markers.RESULT Chest pain(distress) integral scores of treatment group were lower than those of control group(P <0.05),with the total curative rate being 82.50%,notably higher than 52.5% of the control group(P <0.01).The anxiety and depression scores of both groups were evidently reduced after treatment(P <0.01). The comparison of the two groups was P >0.05.TCM syndrome scores of both groups were notably declined(P <0.01).The total curative rate was 75%,which was superior to 20% of the control group(P <0.01).Both electrocardiogram test results and cardiac markers in both groups experienced an evident improvement than before(P <0.05).The comparison of the two groups showed P >0.05.No abnormality was detected in blood,urine,stool or liver and kidney function test in patients of both groups.CONCLUSION Shuangxin Decoction can efficiently improve chest pain,chest distress symptoms as well as TCM clinical syndromes due to coronary heart disease accompanied with psychological disorder.Also,it is helpful for improving de-pression and anxiety,which has been considered as a safe and effective formula for treating these two disorders,worthy of clin-ical application and promotion.