Objective To observe the effect of paeoniflorin on insulin resistance in rats fed with high-fat diet and to investigate the possible mechanisms.Methods Male Sprague Dawley (SD) rats were randomized into 4 groups:normal control,high-fat diet,high-dosage paeoniflorin (HDP group),and lowdosage paeoniflorin (LDP group).The control group was fed with ordinary diet,while the others with highfat diet,paeoniflorin intervention groups were given low-or high-dosage paeoniflorin by intraperitoneal injection.After 6 weeks,fasting serum triacylglycerol (TG),total cholesterol (TC),free fatty acids (FFA),fasting blood glucose (FBG),and insulin were determined.Insulin sensitivity index (ISI) were calculated and then the animals were sacrificed to acquire epididymal fat mass.The tumor necrosis factor alpha (TNFα) and glucose transporter 4 (Glut4) expressions in adipose tissue were detected by quantitative realtime polymerase chain reaction(PCR).Results Compared with high-fat fed group,HDP group had lower epididymal fat pad weight,reduced level of FBG,insulin and FFA (P ＜0.05) and improved ISI(-5.84 ± 0.24 vs-6.44 ± 0.25,P ＜ 0.05).LDP group had similar trends.In adipose tissue,the TNFα expression in LDP and HDP group was lower,Glut4 expression in HDP group was higher than that of high-fat fed group (P ＜ 0.05).Conclusion Paeoniflorin can reduce visceral adipose content,inhibit TNFα expression and increase Glut4 expression in adipose tissue,eventually lower glucose,and improve insulin resistance caused by high-fat diet.

Objective To study the differences in intestinal flora of normal and type 2 diabetic mice, the effect of alcoholic extract of Coreopsis tinctoria Nutt on mouse intestinal flora, and explore the possible relationship between alcoholic extract of Coreopsis tinctoria Nutt, intestinal flora and type 2 diabetes mellitus (T2DM) in mice.Methods Stool samples were collected from the normal control group (A), high dose (1.8 g/kg) (B) and moderate dose (1.2 g/kg) (C) alco-holic extract of Coreopsis tinctoria Nutt model groups, metformin (0.2 g/kg) treatment group (D) and blank control (E) group.16S rDNA real-time quantitative PCR assay was used to determine the levels of Clostridium coccoides and Bacteroides thetaiotaomicron in the stool samples.Pearson analysis method was used to analyze the correlation between the levels of tar-get bacterial species and the fasting blood glucose ( FBG) in the mice.Results 1.Compared with the normal control group, the levels of Clostridium coccoides and Bacteroides thetaiotaomicron in the T2DM model group were significantly low-ered (P=0.017, P=0.002).2.Compared with the model group, the levels of Clostridium coccoides and Bacteroides the-taiotaomicron of the high dose Coreopsis tinctoria Nutt alcohol extract group were significantly different ( 2 weeks: P =0.027, P=0.006;4 weeks:P=0.007, P=0.012).3.The levels of Clostridium coccoides and Bacteroides thetaiotaomi-cron were positively correlated with the FBG level in the mice.Conclusions The alcohol extract of Coreopsis tinctoria Nutt has certain effect on the intestinal flora in type 2 diabetic mice and there is certain correlation between the effect of alcohol extract of Coreopsis tinctoria Nutt and their blood glucose level.

Objective To research the clinical department distribution and drug resistance of Enterobacteriaceae bacteria to pro‐vide a theoretical basis for clinical rational use of antimicrobial drugs .Methods The Whonet 5 .6 software was adopted to conduct the retrospective analysis on Enterobacteriaceae bacteria isolated in the General Hospital of Ningxia Medical University and the Af‐filiated Cardiovascular Disease Hospital from 2011 to 2013 .Results A total of 7 315 strains of Enterobacteriaceae bacteria were i‐solated ;the top three of bacteria were 2 971 strains (40 .6% ) of Escherichia coli ,2 339 strains (32 .0% ) of K lebsiella pneumoniae and 1 117 strains (15 .3% ) of Enterobacter cloacae ;in the source of specimen ,the respiratory tract specimens had the highest isola‐tion rate (46 .6% ,3 410 strains) ,followed by the pus and secretion specimens (13 .9% ,1 015 strains) ,and the urine specimens (13 .0% ,953 strains) ;the isolated bacterial strains were mainly derived from the pediatric department (17 .5% ,1 282 isolates) ,res‐piration department (7 .1% ,518 strains) and ICU (6 .4% ,468 strains) ;the highest sensitivity of antibacterial drugs were carbapen‐ems ,amikacin and piperacillin/tazobactam also maintained a good antibacterial activity ,the resistance rate was 1 .3% - 7 .6% .Con‐clusion Enterobacteriaceae has a higher isolation rate in the clinical specimens and its resistance rates to antibacterial drugs are generally higher .The surveillance on bacterial drug resistance should be strengthened so as to provide a theoretical basis for the ra‐tional use of antimicrobial drugs and effective control of nosocomial infections .

Objective To investigate the safety and tolerability of darbepoetin alfa.a long-lasting erythmpoietin,single intravenous administration in maintenance hemodialysis(MHD) patients. Methods A single center,open clinical trial was carried out.Forty-three stable MHD patients were divided into 5 groups and received darbepoetin α at dosage of 0.225,0.45,0.9,1.8,3.6 μg/kg respectively.The vital signs,symptoms,ECG and laboratory examinations were monitored and detected before and after administration. Results Of the 43 patients (male 26and female 17),the largest tolerable dosage of darbepoetin alfa was 3.6 μg/kg.During the study,the main side effect associated with darbepoetin was hypertension aggravation(7%).One patient died but that was not associated with darbepoetin alfa. Conclusion Darbepoetin alfa is sale and well tolerated.

Objective:To study the correlation between insulin-like growth factor-1 (IGF-1) and its receptor IGF-1R expression in ovarian cancer and clinicopathological features, surgical prognosis.Methods:The positive expression rates of IGF-1 and IGF-1R in ovarian cancer, borderline ovarian tumors and benign ovarian tumors were detected by immunohistochemistry. The correlation of IGF-1 and IGF-1R with clinicopathological features, surgical prognosis was analyzed.Results:The positive expression rate of IGF-1 and IGF-1R in ovarian cancer was 65.63%, 70.31%, significantly higher than 22.22%, 16.67% in borderline ovarian tumors, 5.00%, 10.00% in benign ovarian tumors ( χ2/ P=27.548/0.000, 31.335/0.000) . The mRNA and protein expression of IGF-1 and IGF-1R were also significantly higher than borderline ovarian tumors and benign ovarian tumors ( F/P=31.922/0.000, 26.865/0.000, 34.567/0.000, 27.667/0.000) . The positive expression rates of IGF-1 and IGF-1R in ovarian cancer with FIGOIII-IV stage, tissue differentiation G2-G3 stage, CA125>35 U/mL, Ki-67 positive expression were significantly higher than those in ovarian cancer with FIGOI-II stage, tissue differentiation G1 stage, CA125<35 U/mL, Ki-67 negative expression (IGF-1: χ2/ P=8.505/0.004, 4.980/0.026, 7.481/0.006, 10.907/0.001, IGF-1R: χ2/ P=9.785/0.002, 4.950/0.026, 7.211/0.007, 6.471/0.011) . The total survival time of ovarian cancer patients with positive IGF-1 and IGF-1R was shorter than those of patients with negative IGF-1 and IGF-1R. Conclusion:IGF-1 and IGF-1R are highly expressed in ovarian cancer and are associated with the deterioration of pathological features and surgical prognosis.

Objective:To explore the distribution of integrons in Escherichia coli isolated from community patients with urinary tract infections and their relationship with the phylogenetic groups and antimicrobial resistance. Methods:From November 2015 to December 2018, 152 isolates of E. coli that collected without repetition from the urine samples of outpatients in nephrology of Fengxian District Central Hospital in Shanghai, were studied retrospectively. Bacterial identification and antimicrobial susceptibility analysis was carried out by Phoenix 100 automatic microbiological analyzer. Class 1, 2 integron integrase genes, variable regions of integrons and the phylogenetic groups of isolated E.coli were screened by PCR. The type of promoters and gene cassette arrays of variable regions were determined by sequencing. The relationship of intergon with the phylogenetic groups and antimicrobial resistance was also analyzed. Results:The resistance rate of 152 E. coli to ampicillin was 70.39% (107/152), and the resistance rates to other antibacterial drugs were all less than 40.00%. Among the 152 E. coli isolates, class 1 integron integrase gene intI1 was detected in 65 isolates (42.76%), 8 gene cassette arrays and 14 antimicrobial resistance gene cassettes were detected in 68 class 1 integrons. The most popular gene cassette array was dfrA17-aadA5 (51.47%, 35/68), while the variable regions of class 1 integrons were failed to detected in 12 intI1-positive isolates. Five variable region promoters were detected in 68 class 1 integrons, with the relative weak promoter PcH1 to be the most popular type (77.94%, 53/68). The gene cassette array arr- 2-cmlA5-bla OXA-10-aadA1 was also detected in this study. 65 intI1-positive isolates were mainly belonged to group B2 and D. The class 2 integron integrase gene intI2 was detected in 4 isolates (2.63%,4/152), and their variable region gene cassette arrays were all dfrA1-sat2-aadA1. Conclusions:Class 1 integrons were closely related to antimicrobial resistance in E. coli isolated from community patients with urinary tract infection. Most of the variable region promoters of class 1 integrons were relatively weak promoters. The distribution of each phylogenetic group in the intI1-positive isolates was consistent with the distribution of the overall isolates. The gene cassette array arr-2-cmlA5-bla OXA-10-aadA1 was detected in E. coli.

Objective To investigate the distribution of integrons in clinical isolates of carbapen-em-resistant Acinetobacter baumannii and their relationships to bacterial resistance to antimicrobial agents.Methods A total of 115 carbapenem-resistant Acinetobacter baumannii strains were isolated from clinical samples of patients from January to October, 2017. Phoenix 100 automatic microbiological analyzer was used for antimicrobial sensitivity analysis. Classes 1 and 2 integrase genes and carbapenemase-encoding genes, bla IMP , blaVIM , blaKPC , blaNDM and blaOXA-23 , were screened by PCR. The variable regions of integrons were amplified by long fragment PCR. The types of promoters and gene cassette arrays of variable regions were de-termined by sequencing and overlap PCR. Relationships between integrons and antimicrobial resistance were analyzed. Results The 115 isolates of carbapenem-resistant Acinetobacter baumannii were resistant to most commonly used antimicrobial agents, but sensitive to polymyxin E. All of the isolates carried blaOXA-23 gene and none of them were positive for blaIMP , blaVIM , blaKPC or blaNDM gene. Class 1 integrase gene intI1 was de-tected in 40 isolates (34. 8％ ), while class 2 integrase gene intI2 was not detected. Two gene cassette ar-rays of variable regions, aacA4-catB8-aadA1 (39 isolates) and aacC1-gacP-gacQ-aadA1a (23 isolates), were detected in intI1-positive isolates. Twenty-two isolates carried both aacA4-catB8-aadA1 and aacC1-gacP-gacQ-aadA1a. The upstream promoters of the variable regions were relatively strong promoters, PcH2 and PcS. The gene cassettes of the variable regions endowed bacteria with resistance to chloramphenicol and aminoglycoside antibiotics. The resistance rate of class 1 integron-positive isolates to compound sulfamethox-azole was higher than that of negative strains. However, their resistance rate to ampicillin/sulbactam was lower than that of negative strains. Conclusions Antimicrobial resistance in carbapenem-resistant Acineto-bacter baumannii was serious. Carbapenem resistance was associated with blaOXA-23 gene. The types of pro-moters of variable regions in class 1 integrons were all relatively strong promoters. Class 1 integrons were closely related to sulfonamides resistance.

Objective:To describe the status and influencing factors of the empowerment of caregivers after gastrointestinal tumor.Methods:From March 2019 to October 2019, using convenience sampling method, 124 pairs of gastrointestinal tumor patients and caregivers who were hospitalized in the general surgery and anorectal surgery of the department of Shandong Tai′an City Central Hospital and the Second Affiliated Hospital of Shandong First Medical University were selected as the subjects. A general information questionnaire, Main Caregivers′Empowerment Measurement were used as research tools. To investigate the empowerment ability of caregivers of patients with gastrointestinal tumors after operation.Results:The total caregiver′s empowerment score was 168.81 ± 19.49. The scores of each dimension ranged from high to low: relationship with caregivers (3.83 ± 0.43), expectation of care outcomes (3.80 ± 0.43), awareness of caregiving roles (3.70 ± 0.51), caregivers′ subjectivity (3.37 ± 0.60), caregiving beliefs (3.34 ± 0.67), caregiving knowledge and skills (3.27 ± 0.73), personal resources (2.94 ± 0.65), goodwill caregiving (2.85 ± 0.65) , the scruples around (2.75 ± 0.88). Multiple linear regression results showed that, patient gender, whether the disease is first attack, whether the patient is aware of the disease, the caregiver marriage, and the caregiver working status were the influencing factors of the caregiver′s empowerment ability after gastrointestinal tumor operation ( t values were -8.15-8.72, R2=0.62, all P<0.05). Conclusions:There is room for further improvement in the empowerment of caregivers after gastrointestinal tumor surgery, especially in personal resources, goodwill care and scruples around. The results suggest that medical staff should guide caregivers to emancipate their minds, enhance the breadth of empowerment, and promote self care and fast rehabilitation of patients.

Background/Aims: To investigate the autoantibody against fumarate hydratase (FH), which is a specific liver failure-associated antigen (LFAA) and determine whether it can be used as a biomarker to evaluate the prognosis of acute-on-chronic liver failure (ACLF). Methods: An immunoproteomic approach was applied to screen specific LFAAs related to differential prognosis of ACLF (n=60). Enzyme-linked immunosorbent assay (ELISA) technology was employed for the validation of the frequency and titer of autoantibodies against FH in ACLF patients with different prognoses (n=82). Moreover, we clarified the expression of autoantibodies against FH in patients with chronic hepatitis B (n=60) and hepatitis B virus-related liver cirrhosis (n=60). The dynamic changes in the titers of autoantibodies against FH were analyzed by sample collection at multiple time points during the clinical course of eight ACLF patients with different prognoses. Results: Ultimately, 15 LFAAs were screened and identified by the immunoproteomic approach.Based on ELISA-based verification, anti-FH/Fumarate hydratase protein autoantibody was chosen to verify its expression in ACLF patients. ACLF patients had a much higher anti-FH autoantibody frequency (76.8%) than patients with liver cirrhosis (10%, p=0.000), patients with chronic hepatitis B (6.7%, p=0.022), and normal humans (0%, p=0.000). More importantly, the frequency and titer of anti-FH protein autoantibodies in the serum of ACLF patients with a good prognosis were much higher than that of patients with a poor prognosis (83.9% vs 61.5%, p=0.019; 1.41±0.85 vs 0.94±0.56, p=0.017, respectively). The titer of anti-FH autoantibodies showed dynamic changes in the clinical course of ACLF. Conclusions The anti-FH autoantibody in serum may be a potential biomarker for predicting the prognosis of ACLF.

BACKGROUND AND OBJECTIVEIt has been proven that cytochrome P450 enzyme 2A13 (CYP2A13) played an important role in the association between single nucleotide polymorphisms (SNP) and human diseases. Cytochrome P450 enzymes are a group of isoenzymes, whose sequence homology may interfere with the study for SNP. The aim of this study is to explore the interference on the SNP study of CYP2A13 caused by homologous sequences.

METHODSTaqman probe was applied to detect distribution of rs8192789 sites in 573 subjects, and BLAST method was used to analyze the amplified sequences. Partial sequences of CYP2A13 were emplified by PCR from 60 cases. The emplified sequences were TA cloned and sequenced.

RESULTSFor rs8192789 loci in 573 cases, only 3 cases were TT, while the rest were CT heterozygotes, which was caused by homologous sequences. There are a large number of overlapping peaks in identical sequences of 60 cases, and the SNP of 101 amino acid site reported in the SNP database is not found. The cloned sequences are 247 bp, 235 bp fragments.

CONCLUSIONThe homologous sequences may interfere the study for SNP of CYP2A13, and some SNP may not exist.