Inflammatory caspase-11 senses and is activated by intracellular lipopolysaccharide (LPS) leading to pyroptosis that has critical role in defensing against bacterial infection, whereas its excess activation under pathogenic circumstances may cause various inflammatory diseases. However, there are few known drugs that can control caspase-11 activation. We report here that scutellarin, a flavonoid from Erigeron breviscapus, acted as an inhibitor for caspase-11 activation in macrophages. Scutellarin dose-dependently inhibited intracellular LPS-induced release of caspase-11p26 (indicative of caspase-11 activation) and generation of N-terminal fragment of gasdermin D (GSDMD-NT), leading to reduced pyroptosis. It also suppressed the activation of non-canonical nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome as evidenced by reduced apoptosis-associated speck-like protein containing a CARD (ASC) speck formation and decreased interleukin-1 beta (IL-1β) and caspase-1p10 secretion, whereas the NLRP3-specific inhibitor MCC950 only inhibited IL-1β and caspase-1p10 release and ASC speck formation but not pyroptosis. Scutellarin also suppressed LPS-induced caspase-11 activation and pyroptosis in RAW 264.7 cells lacking ASC expression. Moreover, scutellarin treatment increased Ser/Thr phosphorylation of caspase-11 at protein kinase A (PKA)-specific sites, and its inhibitory action on caspase-11 activation was largely abrogated by PKA inhibitor H89 or by adenylyl cyclase inhibitor MDL12330A. Collectively, our data indicate that scutellarin inhibited caspase-11 activation and pyroptosis in macrophages at least partly via regulating the PKA signaling pathway.

Insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2, also called IMP2) plays an essential role in the development and maturation of germ cells and embryos and is a candidate gene for goat litter size, based on a previous genome-wide selective sweep analysis. In this study, the mRNA expression level of IGF2BP2 was found to be significantly higher in a single-lamb group than in a multi-lamb group. Insertions/deletions (indels) within the goat IGF2BP2 gene, including P4-Ins-13bp and P5-Del-12bp, were verified in 918 Shaanbei White Cashmere (SBWC) female goats. The minor allelic frequencies (MAFs) of P4-Ins-13bp and P5-Del-12bp loci were 0.349 and 0.295, respectively. Analysis using the Chi-square (χ2) test showed that the genotype (χ2=14.479, P=0.006) distribution of P4-Ins-13bp was significantly different between the single-lamb and multi-lamb groups. Correlation analysis demonstrated that P4-Ins-13bp was significantly associated with goat litter size (P=0.022), and individual goats with the homozygous deletion/deletion (DD) genotype produced more litters than other goats. Therefore, considered as a potential molecular marker significantly related to lambing traits, the P4-Ins-13bp mutation of the goat IGF2BP2 gene can be used in goat breeding with practical molecular marker-assisted selection (MAS) to optimize female reproduction and improve economic efficiency in the goat industry.

Objective: To investigate the clinical application value of single nucleotide polymorphism (SNP) haplotype analysis in the preimplantation genetic diagnosis (PGD) of monogenic diseases. Methods: The whole genome amplification products of biopsied trophectoderm cells were analyzed by SNP haplotype analysis and verified by Sanger sequencing. Results: A total of 205 embryos were performed SNP haplotype analysis and Sanger sequencing. Among them, Sanger sequencing failed in 14.63% (30/205) of embryos, and SNP haplotype analysis failed in 0.98% (2/205) of embryos. The failure rate of the latter was significantly lower than that of the former (P＜0.05). There were consistent results in 155 (75.61%) embryos, and inconsistent results in 18 (8.78%) embryos. Forty-five embryos in 41 cycles were performed embryo transplantation. The clinical pregnancy rate was 70.73% (29/41) and the implantation rate was 71.11% (32/45). The results of prenatal diagnosis of amniotic fluid during the second trimester of pregnancy were completely consistent with those of SNP haplotype analysis. Conclusion SNP haplotype analysis is accurate, and its failure rate is lower than that of Sanger sequencing. It can be effectively used in the PGD of clinical monogenic diseases.

OBJECTIVETo demonstrate the clinical applicability of three-dimensional CT angiography by evaluating the anatomic features and variation of inferior mesenteric artery(IMA) and left colic artery(LCA) in order to provide reference to vessel ligation strategy in laparoscopic rectal cancer surgery.

METHODSClinical and image data of 123 patients receiving abdominal multislice CT at The Sixth Affiliated Hospital from 2014 to 2015 were retrospectively analyzed. The images were 3D-reconstructed with computer 3D CT angiography and arterial enhancement phase images were chosen for analysis. Linear distances from IMA root to abdominal aortic bifurcation and from LCA at IMA root level to IMA root were measured. Branch types of IMA, coursing pattern of LCA, and association between LCA and inferior mesenteric vein (IMV) site were summarized.

RESULTSOf 123 cases, 80 were males and 43 were females, mean age was (46.8±16.6) years, body weight was (57.7±10.4) kg, and BMI was (21.3±3.6) kg/m. The average distance from IMA root to abdominal aortic bifurcation was (42.5±7.9) mm, and this distance was closely associated with body weight (OR=4.771, 95%CI: 1.398 to 16.283, P=0.013). Longer distance tended to appear in the heavier patients. LCA and sigmoid artery (SA) originating from same single IMA was found in 61(49.6%) cases; LCA and SA forking at same point in 35(28.5%) cases; LCA and SA coursing together and forking afterwards in 24(19.5%) cases, and LCA disappearing in 3(2.4%) cases. In 71(57.7%) patients, LCA ascended medial to the lateral border of left kidney, while in 16(13.0%) patients, LCA arranged below the inferior border of left kidney. When the LCA site was higher and the distance from LCA to IMA root was closer [distance from LCA to IMA root level was (24.2±9.9) mm, (30.0±15.2) mm and (66.6±12.3) mm, F=83.2, P<0.001]. At the level of IMA root, LCA located medial to IMV in 21(17.1%) cases, located just lateral to IMV in 54(43.9%) cases, and located lateral and ascended far away from IMV in 48(39.0%) cases.

CONCLUSION3D-CT angiography is non-invasive, efficient and accurate in evaluating coursing features and variation of IMA and its branches, which can provide important reference to the surgeons, promising laparoscopic surgery smooth and safe.

Objective:To study the influence of metformin and paclitaxel in the proliferation and apoptosis of human ovarian cancer SKOV3 cells in vitro, and to clarify the synergistic effect of metformin and paclitaxel. Methods:The ovarian cancer SKOV3 cells were divided into blank control group, different concentrations (0.01, 0.50, 1.00, 5.00, 10.00mmol·L-1) of metformin groups and combined treatment groups(metformin combined with paclitaxel with different concentrations).The inhibitory rates of proliferation of SKOV3 cells after treated with different concentrations of metformin were detected by MTT method.The apoptotic rates of SKOV3 cells after treated with different concentrations of metformin were measured by flow cytometry.The inhibitory rates of proliferation of SKOV3 cells after treated with metformin and paclitaxel were determined by MTT method.Results:The MTT results showed that the inhibitory rates of proliferation of SKOV3 cells in different concentrations of metformin groups were increased in concentration-and timedependent manner;there were significant differences compared with blank control group (P<0.05).The flow cytometry results showed that the apoptotic rates of SKOV3 cells in different concentrations of metformin groups were increased;compared with control group, with the increasing of concentrations of metformin, the apoptotic rates of SKOV3 cells in experimental groups 48 h after treatment were increased significantly (P<0.05);the percentages of SKOV3 cells in G0/G1 phase were decreased with the increasing of metformin concentrations(P<0.05) and the percentages of SKOV3 cells in G2/M phase were increased(P<0.05).The MTT results showed that the inhibitory rate of proliferation of SKOV3 cells in 1.00 mmol·L-1 metformin+paclitaxel group was higher than that in 0.50 mmol·L-1 metformin+paclitaxel group was higher than that in 0.50 mmol·L-1 metformin+paclitaxel group(P<0.05),and the inhibitory rates of proliferation of SKOV3 cells in combined treatment groups were higher than those in paclitaxel alone groups(P<0.05).Conclusion:Metformin inhibits the proliferation of ovarian cancer SKOV3 cells through the induction of apoptosis.Metformin can enhance the cell proliferation inhibition of paclitaxel on ovarian cancer SKOV3 cells.The combination of metformin and paclitaxel has a synergistic reaction on SKOV3 cells.

Aim To explore the influence of metformin(a first-line drug for type 2 diabetes) on ATP-induced inflammasome activation and the release of interleukin-1β(IL-1β) by LPS-activated peritoneal macrophages, a commonly-used inflammatory cell model.Methods Peritoneal macrophages were elicited by intraperitoneal injection of 30 g·L-1 thioglycollate into C57BL/6 mice.Inflammasome was activated and cell pyroptosis was induced by LPS plus ATP treatment, and the pyroptotic cells were calculated after propidium iodide(PI) staining.The protein levels of IL-1β and caspase-1 expressed in the cells and released from them into the supernatant were evaluated by Western blot.Immunofluorescent microscopy was recruited to detect the subcellular distribution and fluorescent intensity of the purinergic P2X7 receptor(P2X7R).Results Metformin per se did not induce pyroptosis in LPS-activated peritoneal macrophages, but it significantly and dose-dependently increased cell pyroptosis induced by ATP treatment.At protein levels, maturated IL-1β(17 ku) could not be released from the cells upon single LPS or LPS plus metformin stimulation;but after ATP was added, maturated IL-1β was released into the supernatants of the cells.Moreover, metformin dose-dependently increased the protein levels of both maturated IL-1β and active caspase-1 released by the LPS-activated peritoneal macrophages upon ATP stimulation.Conclusion Metformin intensifies the activation of inflammasome and increases the release of active caspase-1 and maturated IL-1β upon ATP stimulation in the LPS-activated peritoneal macrophages, which should promote inflammatory responses.

Objective:To study the influence of prophylactic bilateral salpingectomy during hysterectomy in ovarian function of the premenopausal women with benign uterine disease and its preventive effect on the pelvic diseases(malignant /benign pelvic lesions or pelvic inflammatory disease).Methods:A total of 100 patients with benign uterine disease underwent hysterectomy were collected and divided into two groups,including 50 patients underwent simultaneous hysterectomy and prophylactic bilateral salpingectomy (prevention group) and 50 patients underwent only hysterectomy (control group).The operative time,blood loss,evacuation time,and hospitalization time of the patients in two groups were detected;the antral follicle count,levels of serum estradiol(E2),follicle stimulating hormone(FSH) and luteinizing hormone(LH)) and the incidence of perimenopausal symptoms of the patients in two groups before operation and six months,1 year after operation were compared;the incidence of pelvic disease of the patients after operation were compared.Results:The operative time,blood loss,evacuation time,and hospitalization time of the patients between two groups were not significantly different(P>0.05).Compared with before operation,the antral follicle count and the E2 levels of the patients 6 months and 1 year after operation in two groups were decreased,and the FSH and LH levels of the patients were inc reased (P<0.01).There were no significant differences in the levels of FSH,LH,E2 and the antral follicle count of the patients between two groups 6 months and 1 year after operation(P>0.05).The incidence of perimenopausal symptoms of the patients in two groups 6 months and 1 year after operation showed no significant difference(P>0.05).The incidence of ovarian malignant tumor and ovarian benign tumor of the patients in two groups 6 months and 1 year after operation had no significantly differences (P>0.05).The incidence of pelvic inflammatory disease of the patients in control group was higher than that in prevention group(P<0.05).There were 2 patients diagnosed as tubal carcinoma in control group 1 year after operation,and the pathological findings showed the atypical cells in 2 patients in prevention group.Conclusion:Prophylactic bilateral salpingectomy during hysterectomy does not damage the ovarian function and can reduce the incidence of pelvic malignant/ benign tumor and pelvic inflammatory disease.

Objective To investigate clinical features of chronic actinic dermatitis (CAD) in Kunming of Yunnan Province.Methods From October 2008 to October 2013,CAD in-patients in our hospital were selected and their general conditions,outcomes of photo-sensitive test and photo-patch test were analyzed in details.Results (1) Among 169 CAD patients,gender ratio (male:female) was 8.9:1.The lesions were mainly distributed on the exposed areas.(2) Photo-sensitive tests of 93 patients showed that the average value of UVA-MPPDwas 7.39 JPcm2 and that of UVB-MED was 20.91 mJPcm2.(3) The results of photo-patch test in 83 patients showed that 48 patients (67.8％) had positive reactions in patch test,and the most common contactant was balsam peru.Fifty-eight patients (69.9％) had positive reactions in photo-patch test indicating that the most common contactant in photo-patch test was balsam peru.Conclusions CAD appears mostly in the middle aged and the aged males and the lesions are mainly distributed on exposed areas.The most common contactants in photo-patch test are balsam peru and perfumed compounds.

Aim To study the mechanism of cucurb-itacin E ( CuE )-induced autophagy in HeLa cells. Methods Improved MTT assay was adopted to meas-ure the effect of CuE on cell proliferation. Western blot was used to determine the phosphorylation levels of downstream signaling proteins of mTORC1 and the ex-pression of autophagy associated proteins. ResultsCuE inhibited the proliferation of HeLa cells in a dose-dependent manner, and the 24-h IC50 of CuE was 4. 01μmol· L-1 . CuE significantly inhibited the phospho-rylation of p70 S6 K in a time-and dose-dependent man-ner as evidenced by decreased phosphorylation levels of
the mTORC1 substrate. Meanwhile, the expression of LC3-II, a marker for autophagosome formation, was elevated by CuE treatment, and was further increased in the presence of chloroquine. Furthermore, CuE re-duced the levels of p62/SQSTM1 . These results indi-cated that CuE induced autophagy in HeLa cells. The decreased levels of phosphorylated ULK1 S757 were posi-tively correlated with autophagy induction in HeLa cells. Conclusion CuE is likely to induce autophagy through inhibiting mTORC1 activity.

Objective To explore the correlation of lung function and IFN-γ and IL-4 levels in peripheral blood in pulmonary tuberculosis patients. Methods The IFN-γ and IL-4 levels in peripheral blood of 75 pulmonary tuberculosis patients(severe 25 cases,moderate 25 casea,and mild 25 cases) and 30 healthy volunteers were measured by ELISA. Meanwhile FEV1 ,FEV1% and MMEF% of lung function in 75 pulmonary tuberculosis patients were measured.Results The levels of IFN-γ[(0. 204 ±0. 018) μg/L] and IL-4[(0. 523 ±0. 035) μg/L] in peripheral blood were significantly different both between tuberculosis group and control group and among tuberculosis patients ( severe,moderate,and mild) (t =7. 685,6. 374 ,all P ＜0. 05) ;FEV1 ,FEV1% ,and MMEF% of severe and moderate tuberculosis patients were significantly lower than those of mild tuberculosis patients and normal reference value; In tuberculosis patients, FEV1% and MMEF% were negatively related with IL-4 level in peripheral blood( r = -0. 46, -0. 43, all P ＜ 0. 05 ), also significantly positively related with the IFN-γ level in peripheral blood ( r = 0. 47,0. 45, all P ＜0. 05). Conclusion Pulmonary tuberculosis morbility may be due to cellular levels of patients. The IFN-γ and IL-4 levels in peripheral blood could affect the lung function of pulmonary tuberculosis patients.