Objective To study plasmid-mediated transfer,plasmid replicon typing,and genetic environment of blaNDM-1 gene in Enterobacteraerogenes(E.aerogenes).Methods E.aerogenes HN-NDM0711 was used as the subject of this research,the transferable properties of plasmid were analyzed by conjugation testing,conjugant was performed stability testing,plasmid type was determined by PCR-based replicon typing (PBRT),downstream and upstream of blaNDM-1 were sequenced using chromosome walking method,genetic context was analyzed by BLASTN and BALSTP,as well as annotated using Vector NTI 11.5.1 software,sequence pipeline graph was made,the sequence was submitted to Genbank through software Banklt.Results The conjugation testing of E.aerogenes pHN-NDM0711 was positive,after positive conjugant was conducted 4-day passage,minimal inhibitory concentrations (MICs) of imipenem and meropenem to all the cloned strains didn't change,blaNDM-1 were all positive.The replicon type was IncA/C;blaNDM-1 gene was localized between ISAba14 and IS91,at upstream of the blaNDM-1,class 1 integron and Tn3 transposon were identified,class 1 integron contained a new mosaic structure of a drug-resistant resistance gene cassette.Conclusion E.aerogenes pHN-NDM071 1,bearing blaNDM-1 gene in IncA/C plasmid,derived from gene recombination under different antimicrobial selection pressure.Antimicrobial use in clinical,industrial and agricultural area should be strictly controlled,so as to reduce the emergence of such bacteria.

Objective To investigate the effects of Shechuangzi San on Th1/Th2 immune function in eczema rats and analyze its mechanisms in treating eczema.Methods Fifty SD rats were randomly divided into blank group,model group,Shechuangzi San high dose group,Shechuangzi San low dose group,and Bingbaiye liquid group,with 10 rats in each group.The rat eczema model was established using 2,4-dinitrochlorobenzene.The blank group and model group were topically treated with 0.9%saline,the Shechuangzi San high dose group was topically treated with a decoction of Shechuangzi San at a concentration of 2.32 g·mL-1,and the low dose group was topically treated with a decoction of Shechuangzi San at a concentration of 1.16 g·mL-1.And the Bingbaiye liquid group was topically treated with the original liquid of Bingbaiye,once in the morning and once in the evening,for a total of 14 days.After the last administration,the skin lesions on the rats'backs were observed and the skin lesion scores were evaluated.The pathological changes of eczema lesions were observed by HE staining,and semi-quantitatively scored.ELISA method was used to detect the levels of IFN-γ,IL-4,IL-18,and IL-33 in rat serum.Flow cytometry was used to determine the Th1/Th2 ratio in the serum.Results Compared to the blank group,the rat skin lesion score in the model group was significantly increased(P<0.05),with a series of skin lesions including edema,erythema,scaling,lichenification appearing on the back.Pathological sections showed excessive keratosis,thickening of cuticular layer,edema of epidermis sponge,infiltration of a large number of inflammatory cells in epidermis,and the semi-quantitative score of skin lesions was significantly increased(P<0.05).Serum levels of IL-4,IL-18,and IL-33 were significantly increased(P<0.05),while the level of IFN-γ was significantly decreased(P<0.05).Serum Th1/Th2 ratio decreased(P<0.05).Compared to the model group,the rat skin lesions in the Shechuangzi San high and low dose groups and the Bingbaiye liquid group were significantly improved(P<0.05),Pathological sections showed that the structure of epidermis was restored to normal,the cuticle became thinner,the spongy edema of epidermis disappeared,epidermal inflammatory cells disappeared,and the semi-quantitative score of skin lesions decreased(P<0.05).Serum levels of IL-4,IL-18,and IL-33 were decreased(P<0.05),while the level of IFN-γ was increased(P<0.05),serum Th1/Th2 ratio increased significantly(P<0.05).and the therapeutic effect of the Shechuangzi San high and low dose groups was superior to that of the Bingbaiye liquid group(P<0.05).Conclusion Shechuangzi San can effectively improve the skin lesions of rats with eczema,repair the pathological damage of skin lesions,and reduce the levels of inflammatory factors.Its mechanism may be related to the inhibition of secretion of inflammatory factors such as IL-4,IL-18,and IL-33,promotion of IFN-γ release,and restoration of Th1/Th2 imbalance.

OBJECTIVETo study the chemical constituents and bioactivity of Teurium pilosum.

METHODVarious column chromatographic techniques were used to isolate the constituents. A combination of EI-MS, NMR spectroscopy and X-Ray were applied to identify the structures. The anti-microorganism was accomplished by disk diffusion method, the antioxidant activity was assayed by the DPPH microanalysis models and the inhibitory activity of alpha-glucosidase was screened In vitro.

RESULTEight compounds were isolated and identified as: glyceryl tristearate (1), 2,5-dioxolanone (2), fernenol (3), stigmasta-5,22-dien-3P-ol (4), 24-nor cholesta-5,22 (E)-dien-3beta-ol (5), ca-spinasterol (6), (3,4-dihydroxyphenyl) acrylate (7), 3,4-dihydroxy phenyl acrylic acid (8).

CONCLUSIONAll compounds have been isolated from the genus for the first time. Compound 3 [IC50 = (37.63 +/- 3.45) mg +/- L(-1)], 6 [IC50 = (178.92 +/- 4.99) mg x L(-1)] and 8 [IC50 = (44.32 +/- 7.02) mg x L(-1)] are of higher inhibitory alpha-glucosidase activity than that of acarbose [IC50 = (1081.27 +/- 12.3) mg x L(-1)]. Compound7 [IC50 = (4.81 +/- 0.96) mg x L(-1)] and 8 [IC50 = (4.16 +/- 0.11) mg L(-1)] showed higher antioxidant activity than that of BHT [IC50 = (35.64 +/- 0.36) mg x L(-1)] and BHA [IC50 = (8.74 +/- 0.39) mg x L(-1)]. Compound 5-8 exhibited inhibitory activity against Fusarium graminearum. Compound 5 and 8 showed inhibitory activity against Botrytis cinerea.

Antifungal Agents ; chemistry ; isolation & purification ; Antioxidants ; chemistry ; Enzyme Inhibitors ; chemistry ; isolation & purification ; Fusarium ; drug effects ; Glycoside Hydrolase Inhibitors ; Plant Extracts ; chemistry ; isolation & purification ; Teucrium ; chemistry ; alpha-Glucosidases ; analysis

OBJECTIVETo explore the differences in sensitivity to rapamycin of five esophageal squamous cell carcinoma cell lines with different differentiation and the changes of sensitivity of the cells after siRNA-interfered expression of p70S6K.

METHODSEffects of rapamycin on proliferation of ESCC cell lines with different differentiation, EC9706, TE-1, Eca109, KYSE790 and KYSE450 cells, were investigated using cell counting kit 8 (CCK-8) assay, and according to the above results, the EC9706 cells non-sensitive to rapamycin were chosen to be transfected with p70S6K-siRNA. The changes in sensitivity of cells to rapamycin were measured in vitro and in vivo using CCK-8 kit, flow cytometry and tumor formation in nude mice.

RESULTSCCK-8 results showed that all the five cell line cells were sensitive to low concentration of rapamycin (<100 nmol/L), but TE-1 and EC9706 cells, which were with poor differentiation, showed resistance to high concentration of rapamycin. After EC9706 cells were treated with 50, 100, 200, 500 and 1 000 nmol/L rapamycin and p70S6K-siRNA, the proliferation rates of EC9706 cells were (48.67 ± 1.68)%, (15.45 ± 1.54)%, (14.00 ± 0.91)%, (10.97 ± 0.72)% and (2.70 ± 0.32)%, respectively, and were significantly lower than that of cells treated with 50, 100, 200, 500 and 1 000 nmol/L rapamycin and control siRNA [(74.53 ± 1.71)%, (68.27 ± 1.35)%, (71.74 ± 2.44)%, (76.23 ± 1.02)% and (80.21 ± 2.77)%] (P<0.05 for all). The results of flow cytometry showed that the ratios of cells in G1 phase of the p70S6K-siRNA, rapamycin and p70S6K-siRNA+ rapamycin groups were (53.82 ± 1.78)%, (57.87 ± 4.01)% and (73.73 ± 3.68)%, respectively, significantly higher than that in the control group (46.09 ± 2.31)% (P<0.05 for all). The results of tumor formation test in vivo showed that the inhibitory effect of rapamycin on tumor growth was stronger after the cells were transfected with p70S6K-siRNA, and the inhibition rate was 96.5%.

CONCLUSIONESCC cells with different differentiation have different sensitivity to rapamycin, and p70S6K-siRNA can improve the sensitivity of cells to rapamycin in vitro and in vivo.

Animals ; Antibiotics, Antineoplastic ; pharmacology ; Carcinoma, Squamous Cell ; drug therapy ; metabolism ; pathology ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Esophageal Neoplasms ; drug therapy ; metabolism ; pathology ; Humans ; Mice ; Mice, Nude ; RNA, Small Interfering ; Ribosomal Protein S6 Kinases, 70-kDa ; genetics ; metabolism ; Signal Transduction ; Sirolimus ; pharmacology ; Transfection

Objective:To investigate the effect of gonadotropin (Gn) on embryo aneuploidy rate and pregnancy outcome during preimplanptation genetic testing for aneuploidy (PGT-A) cycles.Methods:The clinical data of patients undergoing PGT-A cycle at the First Medical Center of the PLA General Hospital from January 1, 2013 to May 31, 2019 were retrospectively analyzed. Patients were divided into younger patient group (<35 years old) and elder patient group (≥35 years old) by maternal age, then divided into two groups in line with Gn dosage (≤2 250 U, >2 250 U), and into four groups by number of oocytes retrieved (1-5, 6-10, 11-15 and ≥16 oocytes). The embryo aneuploidy rate and pregnancy outcome between the groups were compared. Logistic regression was used to analyze the relationship between the cumulative amount of Gn, embryo aneuploidy rate and live-birth rate.Results:A total of 402 cycles (338 patients) and 1 883 embryos were included in the study. (1) In the younger patients, the aneuploidy rate was 52.5% (304/579) in the group of Gn≤2 250 U and 48.6% (188/387) in the group of Gn >2 250 U, with no significant difference between them ( P=0.232). In the elderly patients, the difference in embryo aneuploidy rate between the two Gn group [57.9% (208/359) versus 60.6% (319/526)] was not statistically significant ( P=0.420). (2) The embryonic aneuploidy rate in different protocol of ovary stimulation was analyzed,in the younger group, the embryonic aneuploidy rate in patients using antagonist long protocol was 50.3% (158/314), it was 50.0% (121/242) in agonist long protocol, 52.1% (207/397) in agonist short protocol and 6/13 in luteal phase protocol, no statistical difference was found in above groups ( P=0.923); in the elder group, embryonic aneuploidy rate was 60.8% (191/314) in antagonist protocol, 58.4% (132/226) in agonist long protocol, 59.2%(199/336) in agonist short protocol, 5/9 in luteal phase protocol, respectively,no significant difference was found ( P=0.938). (3) In the younger patients, the aneuploidy rate in 1-5 oocytes group, 6-10 oocytes group, 11-15 oocytes group and ≥16 oocytes group was 37.9% (11/29), 54.0% (94/174), 52.5% (104/198) and 50.1% (283/565) respectively, no significant difference was found between the groups ( P=0.652); while in the elder patients, the difference between aneuploidy rate in each retrieved oocytes group [73.6% (89/121), 57.5% (119/207), 56.3% (108/192), 57.8% (211/365)] was statistically significant ( P=0.046). (4) Logistic regression analysis of age, cumulative dosage of Gn, number of oocytes obtained, and embryo aneuploidy rate showed that there was no association between the amount of Gn and embryo aneuploidy rate ( P>0.05); the increase in maternal age would increase the risk of aneuploidy rate of embryos, which was statistically significant ( OR=1.031, 95 %CI: 1.010-1.054, P=0.004); the increase in oocytes retrived would significantly decrease the risk of aneuploidy ( OR=0.981, 95 %CI: 0.971-0.991, P<0.01). (5) There was no significant difference in biochemical pregnancy rate [55.6% (80/144) versus 52.1% (63/121)], clinical pregnancy rate [50.0% (72/144) versus 47.9% (58/121)] and live-birth rate [46.5% (67/144) versus 40.5% (49/121)] between different Gn dosage groups ( P=0.613, P=0.738, P=0.324). The logistic regression analysis showed that the maternal age, the cumulative dosage of Gn, the number of oocytes obtained, and the ovarian stimulation protocol had no effect on the live-birth rate (all P>0.05). Conclusions:In PGT-A cycle, the dosage of Gn has no association with the embryo aneuploidy rate and pregnancy outcome. In the patients ≥35 years old, the increase in number of oocytes obtained may decrease the risk of aneuploidy. Age is an important factor affecting the embryo aneuploidy in PGT-A cycle.

Objective:To analyze the characteristics of plasmids in KPC-2-producing Serratia marcescens ( S. marcescens) isolates. Methods:Four carbapenem-resistant S. marcescens strains were isolated from four patients admitted to the hepatobiliary ward of Affiliated Cancer Hospital of Zhengzhou University in 2016. BD Phenix-100 was used to identify the strains and detect the minimum inhibitory concentrations (MICs). Homology analysis was performed using pulsed-field gel electrophoresis (PFGE). The modified Hodge test was used to detect the phenotypes of carbapenemase. PCR and gene sequencing were used to detect the types of carbapenem resistance genes. The transferability of plasmids was detected by conjugation test. The characteristics of plasmids were analyzed by genomic alignment method after whole genome sequencing. DNAMAN V9 software was used to compare the amino acid sequences of the replication initiation proteins. A phylogenetic tree was constructed with neighbor-joining method using MEGA7.0. Results:All of the four S. marcescens strains were resistant to carbapenem antibiotics. They were highly homologous according to PFGE. Hodge test results were all positive and the carbapenemase genotype was blaKPC-2. Conjugation test results were positive. The plasmid was a circular DNA of 42 742 bp in length. It had the similar skeleton of incX6 plasmid and the similar amino acid sequence of replication initiation protein. Moreover, it and incX6 plasmid were at the same node in the phylogenetic tree. The blaKPC-2 was located in the core of drug resistance, which was composed of insertion elements including Tn3 family transposons, recombinant enzyme genes, △ISKpn6 and ISKpn27. Conclusions:The plasmid was incX6-like. The blaKPC-2 gene was located in the transposon of △Tn6296. More attention should be paid to the bacteria carrying KPC-2 in incX plasmids.

Humans ; Psoriasis/drug therapy* ; Registries ; China/epidemiology*

OBJECTIVETo evaluate the prognosis and predictive values of preoperative Glasgow prognostic score (GPS) for adenocarcinoma of esophagogastric junction(AEG) patients.

METHODSA retrospective study of 322 AEG patients who received operation between January 2007 and March 2010 in Henan Provincial People's Hospital was performed. Clinical data, pathological characteristics, laboratory parameters and survival data were collected. The GPS was calculated based on C-reactive protein(CRP) and serum albumin(ALB) levels. Univariate and multivariate analysis were used to evaluate the prognostic value of GPS.

RESULTSAmong 322 patients, 0, 1, 2 of GPS were 192, 104 and 26 patients respectively. The median follow-up was 37 (4-73) months. In Kaplan-Meier analysis, median diseases-free survival (DFS) of GPS 0, 1, 2 was 47.0 (95% CI: 31.6-62.4), 15.0 (95% CI: 11.8-8.2) and 4.7 (95% CI: 3.8-5.6) months (P<0.01), and median overall survival (OS) was out of reach, 20.6 (95% CI: 15.8-25.4) and 7.0 (95% CI: 5.8-8.2) months (P<0.01). Univariate and multivariate analysis revealed that GPS was an independent predictor of DFS (P<0.01) and OS (P<0.01) of AEG.

CONCLUSIONGPS is an effective predictor of survival in AEG.

Adenocarcinoma ; C-Reactive Protein ; Disease-Free Survival ; Esophageal Neoplasms ; Esophagogastric Junction ; Humans ; Kaplan-Meier Estimate ; Prognosis ; Retrospective Studies ; Stomach Neoplasms

Objective: To study the predictive and prognostic significance of high-sensitivity modified Glasgow Prognostic Score (HS-mGPS) on the effect of neoadjuvant chemotherapy for advanced gastric cancer. Methods: 117 patients with advanced gastric cancer received neoadjuvant chemotherapy with SOX (oxaliplatin+ S1) or mFOLFOX 6(oxaliplatin+ CF+ 5-FU) regimen. HS-mGPS was calculated according to blood C-reactive protein (CRP) concentration and serum albumin (ALB) level. The correlation between HS-mGPS and clinicopathological characteristics was determined and the predictors of survival were analyzed. Results: 117 patients with stage ⅡB (43 cases), stage Ⅲ (60), and stage Ⅳ (14) received preoperative neoadjuvant chemotherapy. The overall response rate of neoadjuvant chemotherapy was 61.5%(72/117), and the tumor control rate was 88.0% (103/117), with a pathological response rate of 91.5% (107/117). The R0 resection rate was 81.2% (95/117). The median disease-free survival (DFS) was 21.0 (95% CI 6.4-35.6) months. The median overall survival (OS) was 39.0 (95% CI 21.4-56.6) months. Higher HS-mGPS was associated with higher T stage, local lymph-node metastasis, distant metastasis, lower chemotherapy overall response rate and lower pathological response rate (all P<0.05). The univariate analysis and multivariate analysis showed that higher HS-mGPS, presence of local lymph-node metastasis and non R0 resection were associated with poorer DFS and OS (P<0.05). Conclusion HS-mGPS can be used to predict the benefits of neoadjuvant chemotherapy and as an independent prognostic factor for survival in patients with advanced gastric cancer.

This retrospective study aims to demonstrate the effect of antitubercular treatment (ATT) on the pregnancy outcomes and prognoses of patients with genital tuberculosis (GTB) who had received laparoscopy and/or hysteroscopy. This study included 78 patients with infertility and who were diagnosed with GTB through laparoscopy and/or hysteroscopy over the period of November 2005 to October 2015. The recruited patients were divided into ATT and nonATT groups on the basis of ATT duration. The GTB recurrence rates, menstrual patterns, and pregnancy outcomes of the patients were determined at follow-up. Among the 78 patients, 46 received ATT and 32 did not receive ATT. The menstrual volumes of patients in the ATT group significantly decreased relative to those of patients in the nonATT group. GTB did not recur among all patients regardless of treatment. A total of 11 pregnancies (36.7%) in the ATT group and 19 pregnancies (63.3%) in the nonATT group were observed. Pregnancy rates significantly differed (P = 0.002) between the two groups. ATT may decrease the menstrual volume and pregnancy rates of patients who were diagnosed with GTB through laparoscopy and/or hysteroscopy. In addition, ATT did not improve the prognosis of patients with chronic GTB.
Adult ; Antitubercular Agents ; therapeutic use ; China ; Female ; Fertilization ; Humans ; Hysteroscopy ; Infertility, Female ; etiology ; Laparoscopy ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Prognosis ; Retrospective Studies ; Tuberculosis, Female Genital ; complications ; drug therapy ; Young Adult