Objective To explore the influence of Apolipoprotein B(ApoB)gene mutations on the lipid-regulating effect of atorvastatin in patients with hyperlipidemia.Methods 152 patients with hyperlipidemia were selected as the research targets,the ApoB gene polymorphism was detected by polymerase chain reaction-restrictive cut inside fragment length polymorphism(PCR-RFLP)method.All the patients given atorvastatin treatment,and determined lipid levels before and after treatment.Then the lipid level[total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C)]of the different genotypes(no X+carriers and X+carriers)of patients were analyzed.Results The percentage decline of TC and LDL-C of no X+carriers was(28.98±5.21)％,(32.67±7.19)％,respectively,which were higher than those of X+carriers[(18.97±4.01)％ and(26.07±3.45)％](P＜0.05).The percentage decline of TG and increasing of HDL-C of no X+cartiers had no significance difference with those of X+carriers(all P＞0.05).Conclusion ApoB gene mutation influence serum TC and LDL-C levels of hypedipidemia patients,the levels of TC and LDL-C of X+carriers are higher than those of no X+carriers;And the lipid-regulating effect of atorvastatin for X+carriers is more weaker.

Objective To observe the effect of different doses of atorvastatin in the treatment of chronic heart failure of coronary heart disease (CHD), and to provide scientific reference for the choice of clinical medication. Methods 54 cases from June 2015 ~2017 year in April in our hospital with 20mg/d atorvastatin therapy regimen in the treatment of chronic heart failure of coronary heart disease patients as the study group, 54 cases of chronic heart failure of coronary heart disease patients with another force selected by 40mg/d atorvastatin therapy regimen as study group. LDL-C, LVESD, LVEDD, LVEF, hs-CRP, 6MWT, NT-proBNP and other indicators as evaluation basis, through the observation of the two groups before and after treatment the indexes to evaluate clinical efficacy, adverse events were observed after treatment in two groups. Results There was no significant difference between the groups before and after treatment, and there was no statistical significance(P<0.05). The indexes of clinical curative effect were improved in different degree after treatment, and the improvement effect in observation group was better than that in control group (P<0.05). Conclusion Atorvastatin 40mg daily oral drug treatment of chronic heart failure in patients with coronary heart disease is the best choice of atorvastatin dose, the dose range of atorvastatin treatment effectiveness and safety protection, improve clinical symptoms, promote the improvement of the quality of life, worthy of clinical application.

Red-based recombineering has been widely used in Escherichia coli genome modification through electroporating PCR fragments into electrocompetent cells to replace target sequences. Some mutations in the PCR fragments may be brought into the homologous regions near the target. To solve this problem in markeless gene deletion we developed a novel method characterized with two-step recombination and a donor plasmid. First, generated by PCR a linear DNA cassette which comprises a I-Sec I site-containing marker gene and homologous arms was electroporated into cells for marker-substitution deletion of the target sequence. Second, after a donor plasmid carrying the I-Sec I site-containing fusion homologous arm was chemically transformed into the marker-containing cells, the fusion arms and the marker was simultaneously cleaved by I-Sec I endonuclease and the marker-free deletion was stimulated by double-strand break-mediated intermolecular recombination. Eleven nonessential regions in E. coli DH1 genome were sequentially deleted by our method, resulting in a 10.59% reduced genome size. These precise deletions were also verified by PCR sequencing and genome resequencing. Though no change in the growth rate on the minimal medium, we found the genome-reduced strains have some alteration in the acid resistance and for the synthesis of lycopene.
Chromosomes, Bacterial ; genetics ; DNA ; Endonucleases ; metabolism ; Escherichia coli ; genetics ; Genetic Engineering ; methods ; Recombination, Genetic ; Sequence Deletion

Objective To observe the changes of peripheral blood CD27+B cells percentage in patients with HBV-related disease of different severity and the clinical significance.Methods 20 cases of chronic viral hepatitis B,23 cases of HBV-related liver cirrhosis,21 cases of liver cancer were selected,25 cases of healthy controls in the corresponding period who had the physical examination were set as the healthy control group.The peripheral blood in the healthy controls and the patients with HBV-related disease was collected.The cellular immune level changes and CD27+ B cells percentage was detected by flow cytometry,the humoral immunity was detected by immune transmission turbidty method:IgM,IgD.Results (1)Compared with the healthy control group,CD27+B cells percentage significantly decreased in the three groups of patients with HBV-related disease [(5.16 ± 0.36) ％ vs.(4.52 ± 0.22) ％,(2.24 ± 0.15) ％,(0.58 ± 0.02) ％,F =4.32,P ＜ 0.05],and the downward trend gradually obvious as the disease degree exacerbated.nnnnn(2)IgM,IgD in the three groups of HBV related disease patients rised obviously,and the increase range became more obvious as the disease degree exacerbation(F =3.29,5.23,P =0.02,0.03).Conclusion CD27+B cells has a close relationship with HBV-related disease,and rebuilding the body's immune defense system is great importance for evaluating prognosis and the clinical guidance in HBV-related disease.

Objective To compare the pathogenicity between Ureaplasma urealyticum serotype 1 (Up1)and 8 (Uu8) in the genital tract of BALB/c mice.Methods A total of 48 BALB/c mice were randomly and equally divided into four groups:blank control group receiving no treatment,estradiol group pretreated with intramuscular injection of estradiol followed by intravaginal inoculation with sterial liquid culture media,Up1 and Uu8 groups pretreated with intramuscular injection of estradiol followed by intravaginal inoculation with suspensions of Up1 and Uu8 respectively.Three mice were randomly selected from each group to be sacrificed after the collection of vaginal lavage fluid on day 3,7,14 and 21 after the inoculation.Vaginal and uterine tissue specimens were obtained from these sacrificed mice and underwent hematoxylin and eosin (HE) staining.Vaginal lavage fluid samples were subjected to culture of Uu and measurement of tumor necrosis factor-α (TNF-α).Results No evidences were observed for Uu growth in either the blank control group or estradiol group at any of the time points after the inoculation,with the average level of TNF-α in vaginal lavage fluid being (4.17 ± 0.85) pg/ml at these time points in both groups.Uu grew in all the vaginal lavage fluid samples from the Up1 and Uu8 groups at the four time points,with the color change unit (CCU) value decreasing with time.The level of TNF-α in vaginal lavage fluid peaked on day 14 after the inoculation in the Up 1 ((14.93 ± 1.11) pg/ml) and Uu8 ((27.04 ± 24.26) pg/ml) groups.Both Up1 and Uu8 infection caused acute and chronic inflammatory responses in the mice,which were mainly located in the uterus,and Up1 might cause intrauterine adhesion.Conclusions At the same inoculation concentration,no significant difference is found in the pathogenicity between Up1 and Uu8,both of which appear to mainly cause cervicitis.Upl might be partially responsible for intrauterine adhesion in mice.

Objective To establish a method for detection of gene mutations in β-thalassemia by high-resolution melting (HRM) and study its preliminary clinical application.Methods Two common mutations [ IVS-2-654 ( C ＞ T ) and -28 ( A ＞ G ) ]of β-thalassemia in Wenzhou city population were selected.The plasmid DNA fragments of these mutations were constructed by TA clone technology as PCR templates or genotyping controls.A method for detection of β-thalassemia gene mutations based on HRM analysis was established and its specificity,sensitivity and repeatability were methodologically evaluated.One hundred and seventeen patients with clinically suspected β-thalassemia from Second Affiliated Hospital and Yu ying Children's Hospital of Wenzhou Medical College were enrolled into this study.The genomic DNA was extracted from whole blood cells and detected by HRM method.The results were compared with the direct sequencing data.Results HRM method could detect the mutations [ IVS-2-654( C ＞ T) and -28 ( A ＞ G ) ]of β-thalassemia and the results did not show any non-specific amplified fragments.All within-run and between-run coefficients of variation for different DNA types' Tm were smaller than 0.1％.And minimum 103 copies of DNA of each assay and 10％ mutation could be determined by this method.One hundred and seventeen patients with clinically suspected β-thalassemia were detected with HRM and all the results were in accordance with direct DNA sequencing.There were 45 IVS-2-654 ( C ＞ T)heterozygous mutation and 9-28 ( A ＞ G)heterozygous mutation and none homozygous mutation.Conclusion The method of rapid identification of β-thalassemia gene mutations based on HRM analysis is successfully established,which is a convenient,rapid,specific,sensitive and accurate technique for screening gene mutations in β-thalassemia as well as a general technical platform to identify other gene mutations.

OBJECTIVE:To study the hypoglycemic effects of Insulin self-microemulsion for parenteral administration on mod-el rats with type 1 diabetes in vivo. METHODS:Rats were treated with streptozotocin(50 mg/kg)to reproduce model with type 1 diabetes,ip. The model rats were randomly divided into model group (normal saline),positive control group (Insulin injection 2.25 u/kg) and self-microemulsion low,medium and high dose groups (Insulin self-microemulsion 4.5,9 and 18 u/kg);and 10 normal rats were involved in sham-operation group (normal saline). Anesthesia and operation were conducted for all rats. Positive control group was administrated,ip;other rats were parenterally administrated. The blood glucose levels in groups were detected be-fore and after 15-600 min administration. Glucose tolerance test was conducted for the rats in normal control group,model group without glucose,model group with glucose and microemulsion group (Insulin self-microemulsion 9 u/kg). All group were given glucose except model group without glucose. RESULTS:Compared with sham-operation group,the blood glucose levels in model group within 0-240 min were increased,with significant difference(P<0.05). Compared with model group,there was hypoglyce-mic trend in positive control group after 15 min,the blood glucose levels within 30-480 min were decreased,the hypoglycemic peak was 36%,and the peak time was 30 min;there was also hypoglycemic trend in microemulsion low,medium and high dose groups after 30 min,the blood glucose levels within 45-360 min were decreased,hypoglycemic peak was 18%-21%,and the peak time was 90-120 min,with significant differences (P<0.01 or P<0.05). All rats had glucose absorption peak except for model group without glucose in glucose tolerance test,and glucose of rats in microemulsion group reached its peak and then quickly de-creased. CONCLUSIONS:Insulin self-microemulsion can obviously reduce the blood glucose of model rats with type 1 diabetes.

A series of novel riminophenazine derivatives bearing an alkyl substituent attached to N-5 and imino nitrogen at C-3 position of the phenazine ring were obtained through rational drug design, aiming to maintain high anti-tubercular activity, lower toxicity and reduce lipophilicity. All target compounds were prepared by utilizing simple and flexible synthetic route and evaluated against Mycobacterium tuberculosis H37Rv and screened for mammalian cytotoxicity. The results demonstrated that compounds with a cyclopropyl substituent at N-5 position were more active than the reference compound clofazimine. In particular, 2-(4-chloroanilino)-5-cyclopropyl-3-(4-methoxycyclohexyl) imino-3, 5-dihydrophenazine (25) was found to be the most potent compound with low cytotoxicity and lipophilicity. This compound could serve as a valuable lead molecule for further optimization.

Objective:To explore the effect of standardized operation procedure of prone position placement in patients undergoing posterior vertebral pedicle screw system internal fixation.Methods:From May 2019 to May 2021, 98 patients with spinal stenosis who were treated with posterior vertebral pedicle screw system internal fixation in Wuxi Second People's Hospital were selected as the study subject by convenience sampling. The patients were randomly divided into the control group and the observation group with 49 cases in each group. The control group was given routine nursing, and the observation group received standardized operation procedure of prone position placement. The time needed to complete the position placement, the time of position recovery, the incidence of pipeline accidents and the total incidence of pressure injuries were compared between the two groups.Results:After the intervention, the time of position placement and recovery in the observation group were shorter than those in the control group, with statistical differences ( P<0.05) . The incidence of pipeline accidents in the observation group was 2.05%, lower than that in the control group 12.24%; the total incidence of stress injury in the observation group was 6.12%, lower than 18.37% in the control group; but there was no significant difference between the two groups ( P>0.05) . Conclusions:Standardized operation procedure of prone position placement can clarify the division of labor, shorten the prone position placement and recovery time of patients undergoing posterior vertebral pedicle screw system internal fixation, to some extent, it can reduce the risk of pipeline accidents and pressure injuries, and provide safe surgical experience for patients.

Objective:To investigate the professional identity status of nursing students in higher vocational colleges in Shanghai, China, and to analyze the influencing factors.Methods:By cluster sampling, we selected 308 nursing students of grade 2019 from a higher vocational college in Shanghai for a survey with the General Information Questionnaire, Professional Identity Questionnaire for Nurse Students (PIQNS), Stanford Presenteeism Scale-6 (SPS-6), Wong and Law Emotional Intelligence Scale (WLEIS), Workplace Social Capital (WSC), and Nurse Psychological Capital Questionnaire (PCQ). SPSS 22.0 was used for descriptive analysis, the independent samples t-test, one-way analysis of variance, Pearson correlation analysis, and multiple linear regression analysis. Results:The total PIQNS score of the students was (64.93±12.83), the total SPS-6 score was (15.91±4.40), the total WLEIS score was (80.57±15.52), the total WSC score was (32.38±6.33), and the total PCQ score was (95.47±18.63). The PIQNS score was negatively correlated with the SPS-6 score ( r=-0.282, P<0.01), positively correlated with the WLEIS score ( r=0.712, P<0.01), positively correlated with the WSC score ( r=0.659, P<0.01), and positively correlated with the PCQ score ( r=0.681, P<0.01). The multiple linear regression analysis showed that personal interest, emotional intelligence, and psychological capital significantly affected the professional identity of nursing students, entering the regression equation for professional identity. Conclusions:The professional identity of higher vocational nursing students in Shanghai is at a medium level, and personal interest, emotional intelligence, and psychological capital are the main factors influencing professional identity.