OBJECTIVE To investigate the genotype distribution in extended-spectrum ?-lactamases-producing Escherichia coli in our hospital.METHODS Clinical strains of E.coli confirmed to produce ESBLs were collected from our hospital.ESBLs genotype was analyzed by plasmid conjugation,PCR and nucleotide sequencing analysis.RESULTS The susceptiblility rate of ESBLs-producing strains were 100% to imipenem,75.4% to piperacillin/tazobactam.A total of 4 genotypes were identified in the 28 ESBLs-producing strains.CTX-M type was identified in 92.9% of the strains,including CTX-M-14,CTX-M-24 and CTX-M-3.SHV Type was identified in 7.1% of the strains,including SHV-12.CONCLUSIONS ESBLs producers are common in E.coli isolated from our hospital.Most of them are multidrug resistant and CTX-M is the main genotype of ESBLs.Other genotypes of ESBLs are not found.

Objective:To study the expression and clinical relevance of IRAK-M in monocytes in patients with systemic lupus erythematosus. Methods: Real-time quantitative PCR was performed for IRAK-M mRNA measurement and enzyme-linked immunosorbent assay was used for anti-double-stranded DNA antibody ( dsDNA ) and anti-single-stranded DNA antibody ( ssDNA ) . Dynamic scattering turbidimetric immunoassay was applied for complement 3(C3),complement 4(C4) and C-reactive protein(CRP), while Westergren method for erythrocyte sedimentation rate (ESR). Correlation analysis of IRAK-M with SLEDAI,dsDNA,ssDNA,C3, C4,CRP and ESR was computed by Pearson or Spearman. Results:①The result showed that the mRNA expression of IRAK-M in SLE patients was significantly lower than healthy controls (P<0. 05);and the mRNA expression of IRAK-M in active group was significantly lower than stable group ( P<0. 05 ) . ②The levels of dsDNA, ssDNA, CRP and ESR in SLE patients were significantly higher than healthy controls( P<0. 05 );and the levels of C3 and C4 in SLE patients were significantly lower than healthy controls ( P<0. 05 ) . Meanwhile,the levels of dsDNA,ssDNA,CRP and ESR in active group were significantly higher than stable group (P<0. 05),and the levels of C3 and C4 in active group were significantly lower than stable group (P<0. 05).③Correlation analysis showed that the mRNA expression of IRAK-M had a positive correlation with C3 (P<0. 05),and had a negative correlation with SLEDAI,dsDNA,CRP and ESR (P<0. 05),but had no correlation with ssDNA and C4(P>0. 05). Conclusion: This study indicated that IRAK-M play certain significant roles in the pathogenesis of SLE. We can monitor SLE disease activity and prognosis by quantitative detection of mRNA expression of IRAK-M. Meanwhile,it is very necessary to routinely test and regularly monitor the levels of dsDNA,ssDNA,C3,C4,CRP and ESR in SLE.

Objective: To construct siRNA plasmid expression vector in order to knockdown E2F-3 activity. Methods: Sixty-four base-pair oligos for hairpin RNA expression, which targeted E2F-3 gene, were chemically synthesized and annealed. The pRNAT-U6.1/Neo vector was linearized with Bam HI and HindⅢ. Finally, the annealed oligos were inserted into the lined pRNAT-U6.1/Neo to construct RNAi plasmid(pRNAT-U6.1-E2F-3/Neo). The reconstructed RNAi plasmids were i-dentified by electrophoresis after digestion with BamHI and Hind Ⅲ, and were confirmed by sequencing analysis. Results: The recombinant pRNAT-U6.1-E2F-3/Neo vector was identified by polymerase chain reaction, and confirmed by sequencing analysis. The results demonstrated that 64 bp had been inserted into the expected site. Furthermore, the insertion sequence was exactly correct and no mutation site was found. Conclusion: The pRNAT-U6.1-E2F-3/Neo RNAi system was constructed successfully. This will facilitate the study of E2F-3 in bladder cancer cell lines.

Objective: To acquire the expression of E2F3 protein and mRNA in bladder transitional cell carcinoma (BTCC) tissue and normal bladder epithelial tissue, and the relationship between E2F3 expression and the biological behaviors of BTCC thereof. Methods: Immunohistochemistry was used to detect the expression of E2F3 in BTCC(n = 64) and normal bladder mucosa(n = 10). Immunohistochemistry result was analysed by Image-pro Plus software and the expression result was indicated by integrated optical density (IOD). The expression of E2F3 mRNA was investigated using RT-PCR analysis in fresh bladder tumor tissues and normal bladder mucosa. Results: The expression rate of E2F3 in BTCC (32.8%) was higher than that of normal bladder mucosa(P < 0.01). The expression rate of E2F3 was strongly correlated with the pathological grade and clinical stage (P < 0.05;P < 0.01). Immunohistochemistry result indicated that the IOD of E2F3 was significantly higher in BTCC than that of normal bladder mucosa (P < 0.01). The expression level of E2F3 was strongly correlated with pathological grade (P < 0.01). Conclusion: E2F3 was the diagnostic and prognostic index of BTCC, and provided theory basis about the gene target therapy in BTCC.

Objective To explore the potential value of serum lipoxin A4 (LXA4) in monitoring bacterial load and anti-tuberculosis treatment progression in patients with pulmonary tuberculosis (PTB). Methods From January 2021 to January 2022, forty patients with active PTB, who were admitted to Shaanxi Provincial Tuberculosis Prevention and Control Hospital, were selected as the active PTB group, 38 patients with latent tuberculosis infection (LTBI) were selected as the LTBI group, and 28 healthy volunteers who underwent physical examination in our hospital during the same period were included as the healthy control group. The active PTB patients received a 2-month standard anti-tuberculosis chemotherapy, while the other two groups were untreated. Fasting venous blood was drawn from the three groups at enrollment (baseline), after 2 months of treatment, and upon the completion of 6 months of treatment in the active PTB group to measure serum LXA4 levels using enzyme-linked immunosorbent assay (ELISA). The relationship between serum LXA4 level and clinical manifestations, bacterial load, chest imaging manifestations, and treatment progress was analyzed. Results At baseline, serum LXA4 levels in the active PTB group, LTBI group, and healthy control group were [397.72 (210.68, 573.00)], [178.18 (108.17, 271.87)], and [131.06 (76.24, 166.04)] pg/mL, respectively. The levels in the active PTB and LTBI groups were significantly higher than those in the healthy control group, with statistical significance (P<0.01). According to the grading of acid-fast bacilli (AFB) sputum smears at diagnosis, baseline serum LXA4 level increased in the active PTB group with AFB sputum smear grade (P<0.001), and there was a positive correlation between serum LXA4 level and sputum smear grade (rs=0.209, P=0.003). After 6 months of treatment, the serum LXA4 level in the active PTB group was lower than the baseline value (P=0.002). The serum LXA4 level can predict treatment progress, with a baseline sensitivity of 55.0% (22/40), and after 6 months of treatment, 8 patients (20.0%) still showed positive serum LXA4 levels. Conclusions Serum LXA4 may be a useful biomarker for monitoring the progression of PTB treatment.

Objective To explore the inhibiting effect of valsartan and spironolactone on cardiac fibrosis and the expression of integrin ? 1 and fibronectin in the heart of spontaneously hypertensive rats. Methods Eighteen 6 week old SHR were randomly divided into 3 groups with 6 in each: SHR control group, valsartan treating group(30 mg?kg -1 ?d -1 ) and spironolactone treating group ( 20 mg?kg -1 ?d -1 ). Six homogenous male WKY rats served as normal group. After 14 weeks of treatment, systolic blood pressure, left ventricular mass, the ratio of left ventricular mass to body weight (LVM/BW), collagen volume fraction(CVF) and perivascular collagen area(PVCA) were determined and compared among these groups. The expression of integrin ? 1 and fibronectin were also examined by immunohistochemical method. Results Compared with the untreated SHR S, systolic blood pressure was significantly decreased in both treatment groups. LVM/BW〔(2 84?0 14)?10 -3 vs(3 22?0 15)?10 -3 〕, CVF〔(3 21?0 22)%vs(4 00?0 28)%〕, PVCA〔(0 62?0 15)%vs(0 94?0 56)%〕 were lower in both treatment groups, these parameters in SHR V group were even lower than those in SHR S group. Compared with the untreated SHR S, the expression of integrin ? 1 was significantly reduced in SHR V group, while the expression of fibronectin was markedly reduced in both treatment groups. Conclusions Both valsartan and spironolactone could control blood pressure, and effectively inhibit the cardiac fibrosis. Valsartan could also inhibit the expression of cardiac integrin ? 1 and fibronectin, which might be the reason that valsartan is better than spironolactone in inhibiting cardiac fibrosis.

Objective:To investigate the diagnostic values of anti-cyclic citrullinated peptides antibody ( anti-CCP ) and rheumatoid factor( RF) in rheumatoid arthritis( RA) ,and analyse the clinical relevance of prognosis,drug reaction and bone destruction between anti-CCP and RA.Methods: Serum anti-CCP was detected by enzyme-linked immunosorbent assay ( ELISA ) , and RF was detected by immune rate nephelometry.Results:The sensitivity and specificity of anti-CCP in RA were 83.0%and 96.7%,while the sensitivity and specificity of RF in RA were 76.0%and 70.0%.When joint detect anti-CCP and RF,with anti-CCP or RF positive as a positive determination,with anti-CCP and RF negative as a negative judgment,the combined sensitivity was 87.0%,higher than that of detection alone.The combined specificity was 98.3%, higher than that of single detection.There were big different concentrations of anti-CCP among RA patients before treatment, three months after treatment and six months after treatment.There were significant differences between bone erosion and non-bone erosion in RA patients.And the more serious joint damage,the higher the concentrations of anti-CCP.As for treatment,anti-CCP concentrations declined.Conclusion:Combined detection of anti-CCP and RF can significantly improve the diagnosis and differential diagnosis of RA.The concentration of anti-CCP can change with effective treatment,then dynamic monitoring can be used as study drug efficacy.At the same time,the level of anti-CCP in patients with RA can reflect the degree of bone erosion,and serious bone destruction who was poor treatment effect.

AIM:To investigate pharmacokinetics of cinnamon acid and paeonol in Jinkui Shenqi Pill(Radix Rehmanniae Praeparata,Cortex Cinnamomi,Radix Aconiti Lateralis praeparata,Rhizoma Alismatis,etc.)in rabbit.METHODS:RP-HPLC was used to examine blood concentration of cinnamon acid and paeonol in rabbit which was administred Jinkui Shenqi Pill.Chromatographic condition consisted of Symmetry C_ 18(5?m,3.9 mm ?150 mm)chromato graphic column,mobile phase(methanol-1%glacial acetic acid water-solution(45∶55)),flow rate(0.8 mL/min),column temperature(35 ℃),detection wavelength of cinnamon acid(285 nm),detection wave length of paeonol(274 nm).The serum pharmacokinetic parameters were calculated with 3p87 program.RESULTS:Linear range of cinnamon acid ranged from 0.06 ?g/mL to 15 ?g/mL(r=0.999 7),the lowest detectability was 0.054 ?g/mL.Pharmacokinetic process of paeonol in rabbit could be fitted to two-compartment model;Linear range of paeonol ranged from 0.2 ?g/mL to 10 ?g/mL(r=0.999 9),the lowest detectability was 0.02 ?g/mL.Pharmacokinetic process of cinnamon acid in rabbit could be fitted to one-compartment model.CONCLUSION:HPLC detection of serum concentration of cinnamon acid and paeonol is simple,rapid,highly accurate and sensitive.Pharmacokinetic parameters can reveal pharmacokinetics of cinnamon acid and paeonol of Jinkui Shenqi Pill in rabbit.

Objective: To evaluate the daily average intake of dietary fiber of primary school students in Gongshu District, Hangzhou, in order to strengthen the diet guidance and health education. Methods: The method of stratified random cluster sampling was employed to carry out a dietary survey for 887 pupils from 2-6th grade selected from 4 primary schools in Gongshu District, Hangzhou City. The consumption of various kinds of breakfast in pupils’ diet was obtained through dietary survey. The content of dietary fibers in various kinds of food was determined by enzymatic-gravimetric method, and based on this, the daily average intake of dietary fiber of pupils in Gongshu District was calculated. Results: A total of 434 samples of 12 kinds of food in Gongshu District of Hangzhou were determined. The results showed that the dietary fiber content ranged from 0.31 to 2.17 g/100 g in all kinds of breakfast. The difference of dietary fiber content was statistically significant(F=76.50, P=0.00). Among them, noodles and soybean milk were found of more dietary fiber content. However, pupils’ dietary fiber intake for breakfast ranged from 1.82 to 3.04 g/100 g, and the intake was generally low, showing an increasing trend with the increase of grade. The difference of dietary fiber intake among the second to sixth grades was statistically significant (F=18.72, P=0.01). After comparison by SLD method, it was found that the difference of dietary fiber intake between the second and third grade (P=0.22), and between the fifth and sixth grade(P=0.30) were not statistically significant, but the others were statistically significant(P<0.05). Conclusion The unreasonable dietary structure and poor breakfast quality are common phenomena. Well-tailored education programs slaping of adequate and blanced dietary behaviors are in great needs.

Objective To understand the correlation between health beliefs and family environment of stroke patients. Methods A questionnaire survey was carried out on 115 stroke patients with the first onset of stroke by using the special health belief simple table (SF-HBMS) and the family environment scale (Chinese version FES-CV), and the correlation was analyzed. The scores of each subscale of the family environment were compared with the domestic norm. Results The total score of health belief (75.15 ± 10.20) was at the middle level. There were significant differences in age (F=8.41), education level (F=4.44), complications (F=4.05), family history (t=2.68) and first visit time (F=3.76) among different characteristics of health belief scores (P < 0.01 or 0.05). The score of intimacy (6.23 ± 1.27) in family environment, emotional expression score (5.30 ± 1.97), success score (5.88 ±1.62), cultural score (4.54 ± 2.20) and organizational score (5.60 ±1.67) were all lower than the domestic norm and spear. The score of shield score (3.16 ± 2.00) was higher than that of domestic norm (P<0.01 or 0.05), and the total score of health belief was positively correlated with family intimacy (r=0.190), emotional expression (r=0.204), culture (r=0.206) and tissue (r=0.227) (P<0.05), and was negatively correlated with the contradiction (r=-0.186, P<0.05); regression analysis, whether there were family history (β=0.338, P<0.01), first onset time (β=0.242, P<0.01), family intimacy (β=1.614, P<0.05), emotional expression (β=1.114, P<0.05) were the factors affecting the health belief level of first stroke patients. Conclusions The level of health belief is closely related to family environment. It is suggested that the clinical medical staff should pay attention to the negative emotion and family psychological intervention, provide psychological support for the patients and their families, promote the promotion of their health beliefs, and reduce the rate of recurrence and disability.