Objective To establish an infection model using Caenorhabditis elegans (C.elegans)-extensively drug-resistant Klebsiella pneumoniae (XDRKP)system.Methods Clinically isolated XDRKP strains were used to infect C.elegans in the liquid killing assay,the nematode survival and the number of bacteria in C.elegans digestive tract was observed.Results C.elegans was significantly retarded after being infected by XDRKP,different concentra-tions of XDRKP led to different patterns of the worm death.Log-rank test showed that survival curves of C. elegans infected with 1 .5×106 CFU/mL of XDRKP and E.coli OP50 (control)were not significantly different (χ2 =0.08,P >0.05);survival curves of C.elegans infected with 1 .5 ×107 CFU/mL,1 .5 ×108 CFU/mL of XDRKP and E.coli OP50 were significantly different(χ2 =229.37,275.98,respectively,both P <0.001).The survival rates of 1 .5×108 and 1 .5 ×107 CFU/mL XDRKP groups were both lower than that of the control group.Supernatant suspension obtained from test was performed bacterial culture,identification and antimicrobial susceptibility testing, XDRKP was determined.After being infected with XDRKP 4,6,12,and 24 hours,the total number of bacteria in C.elegans were(0.28±0.02)×105 CFU/mL,(0.50 ±0.38)×105 CFU/mL,(1 .73 ±0.56)×105 CFU/mL,and (2.62±0.53)×105 CFU/mL,respectively,the number of bacteria in C.elegans digestive tract was significantly different at different time points (F =1 363.39,P <0.001).Conclusion The infection model of C.elegans-XDRKP is established successfully.

Objective To study the relationship between serum osteocalcin and parameters of glucose and lipid metabolism in elderly men.Methods The bone metabolism index such as serum osteocalcin was measured by electrochemiluminescence immunoas -say in 206 old male patients , incluiding 69 patients with type 2 diabetes mellitus .The parameters of glucose and lipid metabolism were also measured and the correlation between the parameters and serum BGP were analyzed .Results Serum BGP and beta-CTx concen-trations were significantly lower in patients with type 2 diabetes than those in normal glucose tolerance group [ ( 9.57 ±4.74 )μg/L vs (13.22 ±10.35)μg/L, P <0.05;(0.25 ±0.19)μg/L vs (0.35 ±0.29)μg/L, P <0.05].Compared with the group in low level of BGP, fasting blood glucose and HbA1c were reduced in high-level-BGP group[(5.89 ±2.10)mmol/L vs (5.28 ±1.38)mmol/L, P<0.01;6.30%±1.03% vs 5.98%±0.61%, P <0.01].Triglyceride (TG) was positively correlated with serum BGP ( r =0.146, P =0.032).Stepwise multiple regression analysis showed that TG and HbA 1c were independently associated with serum BGP level(β=1.995, P <0.01;β=-1.483, P <0.05).Conclusions Serum TG and HbA1c are independent factors related to serum BGP in elderly men .

Metal or semi-metal elements have numerous physiological and biochemical properties and have correlations with the process of occurrence and development of Zheng and Chinese herbal medicine pharmacodynamics mechanism. This article will expound the current situation of traditional Chinese medicine (TCM) research based on metal elements from the researches of Zheng, medicine theory, pharmacodynamic mechanism, prescription principles, medicine concocted theory and medicine quality control in TCM. In addition to putting forward the shortages of current researches, we also introduce metallomics, the member of Omics in systems biology to offer a new idea for modernization of TCM based on metal elements.

AIM:To explore whether YAP protein is important in induced pluripotent stem cell ( iPSC)-induced cardiovascular progenitor cell and/or vascular smooth muscle differentiation .METHODS:Using episomal vector based reprogramming , we generated human iPSCs from donor fibroblasts .We used both this iPSCs and human H 1 embryonic stem cells to differentiate into vascular smooth muscle cells (VSMCs) through cardiovascular progenitor cells (CVPC).Western blotting, qPCR and immunofluorescence microscopy were used to check the expression of YAP and related genes during this differentiation process .RESULTS:The results showed that iPSCs expressed pluripotent stem cell markers, such as Oct4, Nanog, Sox2, TRA-1-60 and SSEA3, and could form teratoma in SCID mice.YAP was highly expressed in pluripotent stem cells , but dramatically decreased when CVPC differentiation started .YAP gradually increased dur-ing CVPC three-day differentiation.The TAZ and YAP binding partner TEAD1, but not TEAD2 and TEAD4, have similar expression pattern in CVPC differentiation .Immunofluorescence result confirmed that YAP was activated and accumulated in nucleus .Interesting-ly, both YAP and phosphorylated YAP expression decreased to very low level after CVPC differentiated into VSMCs in 7 days.TEAD4 and TAZ also decreased, while TEAD1, TEAD2 and TEAD3 expression did not change during VSMC differentiation .CONCLU-SION:YAP and TEAD1 expression increased during CVPC differentiation , while YAP and TEAD4 expression decreased from CVPC to VSMCs differentiation , which suggested YAP might have different function during diverse cell differentiation .

AIM: To define the gene expression changes of vascular smooth muscle cells (VSMCs) in response to norepinephrine (NE). METHODS: The expression adrenergic receptors (AR) were determined by radioligand binding assay in A7r5 cells. Gene expression profiles were identified by cDNA microarray after A7r5 cells were treated with NE for 24 h, and mRNA expressions of ? 1A -AR and ? 1B -AR were confirmed by real-time PCR. RESULTS: ? 1-AR and ?-AR existed in A7r5 cells. Seventy-five genes with changed expression in response to NE were screened out. These genes are involved in cell structure, cell/organism defense, metabolism, signal transduction and so on. ? 1A -, ? 1B -AR mRNA expression identified by microarray and realtime quantitive PCR displayed similar patterns. CONCLUSIONS: Gene expression profile in response to NE was analyzed comprehensively with the microarray technique. NE induces many kinds of different function genes in A7r5 cells, which may provide a novel insight into the particular role of NE that modulates multiple aspects of biological function in VSMCs. [

Objective To prepare Wurenchun solid dispersion of alcohol extracts of Fructus Schisandrae Chinensis so as to improve the dissolution of its active compound in vitro.Methods The Wurenchun solid dispersions were prepared with various carriers and drug/carrier ratios by mixing the carrier in alcohol extractive solution of FSC directly,and the apparent solubility and dissolution of deoxyschisandrin in them were tested and compared.Results The apparent solubility and dissolution of deoxyschisandrin of Wurenchun solid dispersion(extracts: polyvinylpyrrolidone(PVP) K30=1∶3) were increased remarkably to 5.06 ?g/mL and 43.2% in water individually,including dispersed and dissolved drug whose particle size is below 0.22 ?m,compared with that of the self-prepared Wurenchun capsules.Conclusion Wurenchun solid dispersion made of PVP K30 can remarkably enhance apparent solubility and dissolution of the active compound in vitro.

In order to explore a potential indicator of predicting the occurrence and development of gestational trophoblastic tumor, the expression of c-erbB2 oncogene in human normal placenta, hydatidiform mole and choriocarcinoma was investigated. The expression of c-erbB2 was detected immunohistochemically by monoclonal antibody against the gene on the formalin-fixed paraffin sections of 21 hydatidiform moles, 21 invasive moles, 20 choriocarcinomas and 30 normal placentas. Results showed that the expression level of c-erbB2 was significantly higher in gestational trophoblastic tumor than in hydatidiform mole and normal placenta of midterm and term pregnancy (P < 0.05), while there was no significant difference between patients with gestational trophoblastic tumor of stage III, IV and those of stage I, II. It was demonstrated that overexpression of c-erbB2 may closely associated with malignant transformation of hydatidiform mole, not only providing important insight into pathogenesis of gestational trophoblastic tumor, but also having an important significance for the early diagnosis and early treatment of gestational trophoblastic tumor.
Biomarkers, Tumor ; Choriocarcinoma ; metabolism ; Female ; Genes, erbB-2 ; Humans ; Hydatidiform Mole ; metabolism ; Hydatidiform Mole, Invasive ; metabolism ; Placenta ; metabolism ; Pregnancy ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Uterine Neoplasms ; metabolism

To investigate the relationship between the insulin resistance (IR) of polycystic ovarian syndrome (PCOS) rat model induced by dehydroeplandrosterone (DHEA) and hormonal changes in the ovarium and the resistin mRNA levels in adipose tissue, 21-day-old female SD rats were divided into two groups in pairs. The rats in group 1 were injected daily (s.c.) with DHEA for up to 20 days and the rats in group 2 injected with oil at the same time. Ovarian weight, serum insulin levels and sex hormone levels in rat blood of both groups were determined. Oral glucose tolerance tests, light microscopic and electronic microscopic examination were performed. The levels of resistin mRNA in adipose tissue were measured by reverse transcription-polymerase chain reaction (RT-PCR). Our results showed that the ovarian weight in group 1 was greater than that in group 2 (P<0.05). The ovaria in group 1 showed multiple follicular cysts, The serum testeosterone and etrasdiol concentration were significantly higher in group 1 than those in group 2 (P<0.001 and P<0.05 respectively), so as the fasting serum glucose (P<0.001) and fasting serum insulin (P<0.05). The value of 1/FINS x FGC was significantly higher in group 1 than that in group 2 (P<0.001). Moreover, the resistin mRNA level of white adipose tissue in the DHEA-induced group was significantly higher than that in the control rats (P<0.05). It is concluded that the DHEA-induced PCOS rat models were similar to those of the patients with PCOS, and the IR was observed. Resistin secreted by adipose tissue may mediate IR in PCOS, and it is likely involved in the pathogenesis and development of PCOS.
Adipose Tissue ; metabolism ; Animals ; Animals, Newborn ; Dehydroepiandrosterone ; Estradiol ; blood ; Female ; Insulin Resistance ; Polycystic Ovary Syndrome ; chemically induced ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Resistin ; biosynthesis ; genetics ; Testosterone ; blood

OBJECTIVETo investigate the effect of Ca(2+) on lipopolysaccharide (LPS)-induced NF-kappa B activation in pancreatic acinar cells and the role of NF-kappa B in LPS-induced acinar cell injury.

METHODSMale rat pancreatic acinar cells were isolated by collagenase digestion, then exposed to varying concentrations of LPS (from 1 to 20 mg/L) in the presence or absence of EGTA. At various time points (30 minutes, 1 hour, 2 hours, 4 hours and 10 hours) after treatment with the agents, cell viability was determined by MTT. Nuclear translocation of NF-kappa B's subunit p65 was visualized by immunofluorescence staining and nuclei protein was extracted to perform EMSA which was used to assay the activity of NF-kappa B binding to the DNA sequence containing the recognition site of NF-kappa B.

RESULTSLPS induced cell damage in a time- and concentration-dependent manner while EGTA attenuated LPS-induced cell damage (P < 0.05). NF-kappa B p65 immunofluorescence staining had increased intensity in the cytoplasm and indicated that nuclear translocation occurred within 30 minutes and its zenith was reached at 1 hour after LPS (10 mg/L) treatment. Testing of NF-kappa B DNA binding activity showed the same alteration phase as p65 immunofluorescence staining. NF-kappa B activation preceded the pathological alteration of pancreatic acinar cells. The Ca(2+) chelator EGTA inhibited LPS-induced NF-kappa B activation.

CONCLUSIONSNF-kappa B activation is an important early event in LPS-induced injury to pancreatic acinar cells. Ca(2+) is an important mediator in the process of LPS-induced NF-kappa B activation.

Animals ; Calcium ; pharmacology ; Lipopolysaccharides ; pharmacology ; Male ; NF-kappa B ; physiology ; Pancreas ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley

Objective To study the efficiency and accuracy of artificial intelligence (AI) system based on fundus photograph in diabetic retinopathy(DR)screening,and evaluate the clinical application value of AI system. Methods A diagnostic trial was adopted. Total of 13683 color fundus photos were collected in Zhaoqing Gaoyao People's Hospital from March,2017 to November,2018. The AI system for DR (ZOC-DR-V1) was established,based on transfer learning + NASNet algorithm,by training 4465 precisely labeled fundus images (2510 normal,and 1955 with any stage of DR). One thousand confirmed fundus images (300 normal and 700 with any stage of DR),diagnosed by AI ( AI group ) and doctors ( 3 ophthalmologist doctors and 3 endocrinologist doctors ) ( doctor group ) , respectively. Ophthalmologist group and endocrinologist group were both composed of primary,intermediate and senior physicians. The mean reading time of each image and the total time of 1000 images were recorded. The accuracy and efficiency of AI system and doctor groups were compared. The reading process was divided into two stages. The diagnostic coincidence rate and the average reading time of each group between the two parts were calculated and compared. This study protocol was approved by Ethic Committee of Zhongshan Ophthalmic Center, Sun Yat-sen University (No. 2017KYPJ104). Results After training,the diagnostic coincidence rate of AI system (ZOC-DR-V1) in test set was 94. 7％,AUC was 0. 994. In this "man-machine to war",the diagnostic coincidence rate of primary,intermediate and senior endocrinologist was 94. 0％,91. 4％ and 93. 4％;the diagnostic coincidence rate of primary,intermediate and senior ophthalmologist was 92. 7％,94. 4％ and 95. 6％;the diagnostic coincidence rate of AI system was 95. 2％. There was no difference in the diagnostic coincidence rate between AI system and senior ophthalmologist ( P = 0. 749 ) . The mean reading time of each image of primary, intermediate and senior endocrinologists was (4. 63±1. 87),(3. 74±3. 47) and (5. 71±3. 47) seconds,and the total time of 1000 images of primary,intermediate and senior endocrinologists was 1. 29,1. 04 and 1. 58 hours;the mean reading time of each image of primary,intermediate and senior ophthalmologists was ( 7. 25 ± 6. 58 ) , ( 5. 18 ± 5. 01 ) and ( 5. 18 ± 3. 47 ) seconds,and the total time of 1000 images of primary,intermediate and senior endocrinologists was 2. 02,1. 44 and 1. 44 hours;the mean and total time of AI system was (1. 62±0. 67) seconds and 0. 45 hours,and the reading time of AI system was significantly shorter than that of the doctor groups (all at P=0. 000). The diagnostic coincidence rates between previous and posterior part of primary endocrinologist, primary and intermediate ophthalmologist were significantly different (χ2=11. 986,6. 517,10. 896;all at P<0. 05),and the mean reading time in the posterior part was significantly shorter than that in the previous part of intermediate and senior endocrinologist and primary ophthalmologist (t=4. 175,8. 189,5. 160;all at P<0. 01). While the reading time of AI system remained stable throughout the process(χ2=3. 151,P=0. 103;t=0. 038,P=0. 970). Conclusions The ophthalmic AI system based on fundus images has a good diagnostic efficiency,and its diagnostic coincidence rate can compare with senior ophthalmologist,providing a new method and platform for large-scale DR screening.