In order to obtain the serotype distribution of E.coli from duck and to screen the vaccine bacterial strains,the serotype identifications and biological characteristics of E.coli were analyzed in recent years from Shandong,Hebei and other areas of commercial duck field;selections of vaccine strains were detected by the virulence and immunogenicity.Totally 44 isolated bacterial strains of E.coli from duck were identified to a total of six serotypes:O78,O93,O76,O2,O92 and O32.The O78 serotype was the dominant serotype,accounting for 56.8％ (25/44);O93 serotype for 15.9％ (7/44) according to bacterial Oantigen typing.The strain SD (O78 serotype) was confirmed to have strong virulence and good immunogenicity.The O78,O93 and O76 are the dominant serotypes of duck E.coli in the study areas.The SD strain could be used as the candidate for the next development of inactivated vaccine.

Objective:To study the antidepressant effect of Baihe Zhimu decoction (BZD)and its influence in the key factors (CaM,CaMKⅡ,CREB)of CaM signaling pathway in hippocampus of the rats with depression,and to explore the antidepressant effect of BZD. Methods:Fifty rats were divided into control group,model group, fluoxetine group,low and high doses of BDZ groups (n = 10).Expect for control group,all the rats in other groups were made depression models by means of chronic unpredictable mild stress along with isolated raising,for 21 d.Then the rats were fed with NS, fluoxetine (1.8 mg · kg-1 ), and BZD (1.5 and 3.0 g · kg-1 ), respectively;for 28 d.The learning and memory ability,autonomous activities and the fixed time in 5 min of the rats were tested by Morris water amaze,Open-field Test and Forced Swimming Test respectively. The damage and repair status of hippocampal neurons were observed by Nissl staining method;the expression levels of CaM,CaMKⅡ protein,CREB mRNA in hippocampus of the rats were detected by Western blotting and RT-PCR method. Results:Compared with model group,the total time of rats in the platform quadrant of Morris water maze in BZD groups and fluoxetine group,the total distance and the number of crossing platform were increased (P <0.05 or P <0.01),and the time of first crossing platform were shortened (P <0.01);the total scores in open field test were increased (P <0.01),the fixed time with 5 min in the forced swimming test was shortened (P <0.05 or P <0.01).Compared with fluoxetine group,the fixed time within 5 min of the rats in swimming test was shortened (P <0.05).The result of Nissl staining showed that the hippocampal neuron injury in BZD groups and fluoxetine group was improved compared with model group.The molecular test results showed that the CaM and CaMKⅡprotein expression levels in hippocampus of the rats in BZD groups and fluoxetine group were increased compared with model group (P < 0.05 or P < 0.01).Compared with model group,the CREB mRNA expression levels in fluoxetime group and BZD groups were increased (P < 0.05 or P < 0.01).Conclusion:BZD has antidepressant effect and can improve the hippocampal neuron injury of the rats with depression and its mechanism is related to increasing the expression levels of CaM,CaMKⅡ and CREB in hippocampus CAM signaling pathway of the rats.

Objective To investigate Nuclear factor-κB (NF-κB) expression in tumor associated inflammation in patients with adamantinomatous craniopharyngima..Methods Fifty-four patients (31 male and 23 female) with craniopharyngioma from 3 to 66 years of age were recruited from May 2004 to March 2006.NF-κB and Osteopontin (OPN) expression in human craniopharyngiomas were detected using immunohistochemical staining.High-sensitivity C-reactive protein (hs-CRP), a systemic marker of inflammation, was examined in patients' tumor hydatid fluid, cerebrospinal fluid and serum.Results NF-κB expression was significantly increased in the adamantinomatous craniopharyngioma.Spearman;s correlation analysis demonstrated that NF-κB expression was associated with OPN expression.The hs-CRP level was also increased in the tumor hydatid fluid (4.28±0.90 mg/mL), cerebrospinal fluid (0.035±0.006 mg/mL) and serum (1.72±0.54 mg/mL) in patients with adamantinomatous craniopharyngioma.Conclusions NF-kappa B is closely associated with tumor associated inflammation which further mediates adhesion of tumor to surrounding important structures in patients with adamantinomatous craniopharyngioma.

Objective To construct,express,purify and identify the gene encodi ng major outer membrane protein of Neisseria gonorrhoeae (Porin I, or PI). Metho ds The gene encoding for PI of N.gonorrhoeae was amplified by PCR and cloned int o expression plasmid pGEX-4T-2 to form pGEX-4T-2/PI recombinants. A high lev el expression of GST-PI fusion protein was obtained in GST gene fusion system (GST:glutathione S transferase). The analysis indicated that the expressed pr otein was present predominantly in the insoluble form. Therefore, the induced pr otein was purified by SDS-PAGE, and bands corresponding to polypeptides of GST-PI fusion protein were excised and subjected to electroelution. A dot immunoch romatographic assay was employed to demonstrate whether the purified protein was gonococcal PI specific. Results The pGEX-4T-2/PI expression recombinants were constructed,expressed,purified and identified successfully. SDS-PAGE analysis and dot immunochromatographic assay suggested that the recombinant GST-PI fusio n protein was a 60 000 molecular weight protein andidentical in size to native PI and reacted with anti-PI monoclonal antibody. Conclusion Our results may lead to a potentiality for further study of diagnosti c kits and vaccine for Neisseria gonorrhoeae.

Objective To retrospectively evaluate and analyze the clinical effect of posterior pedicle subtraction osteotomy in treating chronic, posttraumatic thoracolumbar kyphosis. Methods Nineteen patients (11 males and 8 females) with chronic, posttraumatic thoracolumbar kyphosis were corrected surgically. The patients were at age range of 29-61 years (mean 42 years). Preoperative kyphosis Cobb angle ranged from 31° to 63° (mean 47°) and trauma history ranged from 8 months to 63 months (mean 29 months). All patients were treated with pedicle subtraction osteotomy according to the size of Cobb angle, extent of spinal stenosis and source of compression. Results Sagittal alignment was improved to average 40.2°, with a correction rate of 85.8%. Two patients developed postoperative leakage of cerebrospinal fluid. Among them, one was combined with encephalic infection and cured with active treatment, and the other developed postoperative wound infection, which were treated conservatively with antibiotics and local wound care. There were no other severe complications. The average follow-up period was 15 months (range 6-41 months). At the last follow-up, clinical symptoms and neurological function were improved significantly. Neither loss of correction nor failure of internal fixators was observed. X-ray and dynamic X-ray films showed a 100% fusion in all patients. Conclusions The single-stage posterior pedicle subtraction osteotomy is a safe and effective procedure for correction of posttraumatic thoracolumbar kyphosis. It is possible and safe to obtain a correction within 55° on single segment by this technique.

Objective To observe the initial mechanism investigation in immune escaping correlated T cell proportion of Lewis lung cancer by Fuzheng-Peiyuan formula. To provide a theoretical basis for the application of the Chinese medicine of strengthening the body resistance in clinics. Methods The healthy C57BL/6J mice were divided into six groups by the random number table: the Chinese medicine group, the Chinese medicine plus chemotherapy group, the chemotherapy group, the model group and the normal control group. There were 6 mice in each group. All the groups expect the normal control group were inoculated subcutaneously with Lewis lung cancer cell suspension 0.2 ml. The medicine given to the Chinese medicine group were orally Fuzheng-Peiyuan formula with 8.17 g/kg. The combination group was given intraperitoneal injection of cyclophosphamide 60 mg/kg and orally Fuzheng-Peiyuan formula 8.17 g/kg. The chemotherapy group received the injection of cyclophosphamide 60 mg/kg and intragastric administration of normal saline. The model group and normal control group were administered with saline. After continuous administration for 13 days, the expressions of CD4, Foxp3, CD8 and CD28 in spleen cells of tumor bearing mice were observed by flow cytometry. Results Compared with the model group, the chemotherapy group and the chemotherapy combined with Chinese traditional medicine group showed that tumor weight (1.76 ± 0.42 g, 1.40 ± 0.43 g vs. 4.37 ± 0.59 g) significantly decreased (P<0.01); and the CD4+ Foxp3+ cells of mouse spleen cells in the Chinese medicine group and Chinese medicine plus chemotherapy group (11.25% ± 1.69%, 9.30% ± 2.68% vs. 14.21% ± 1.50 %) significantly decreased (P<0.05 or P<0.01). Conclusions The result initial proved that the Chinese medicine could strengthen the body resistance, adjust the proportion of Treg and CTL in spleen T cell in tumor-bearing mouse.

Objective To study the effect of olanzapine combined with repetitive transcranial magnetic stimulation(rTMS) in the treatment of phonism dominated schizophrenia.Methods From August 2015 to November 2016,112 patients with phonism based schizophrenia in Ningbo Kangning Hospital were selected in the research . According to the different treatment ,the patients were divided into observation group and control group ,with 56 cases in each group.The observation group was treated with olanzapine combined with rTMS chemotherapy ,the control group was treated with olanzapine.Before and after treatment,the positive and negative symptom scale ( PANSS) score, Wisconsin Card Sorting Test ( WCST) score of the two groups were observed.The clinical efficacy was compared between the two groups.Results Before treatment,there were no statistically significant differences in PANSS score and WCST score between the two groups ( all P＞0.05).After treatment for 1 week,2 weeks,4 weeks,the PANSS scores of the two groups were significantly lower than those before treatment (F＝170.710,106.028,28.530,30.328, 25.806,10.832,203.342,372.253,all P＜0.05).The PANSS scores decreased more significantly in the observation group.After treatment for 1,2,4 weeks,the scores of positive symptoms in the observation group were (25.95 ±3.50)points, (24.72 ±4.50)points and(16.51 ±2.70)points,respectively,which were significantly lower than those in the control group[(27.27 ±2.03)points,(27.80 ±5.37)points,(19.53 ±3.07)points](t＝2.441,3.290,5.528,all P＜0.05).After treatment for 1 week,2 weeks,4 weeks,the continuous response scores in the observation group were (45.62 ±5.41)points,(44.69 ±4.91) points,(35.89 ±3.30) points,respectively,which were significantly lower than those in the control group[(50.61 ±5.35)points,(46.80 ±5.14)points,(42.70 ±5.04)points](t＝4.908, 2.221,8.459,all P＜0.05).The scores of continuous errors in the observation group were (49.47 ±4.59) points, (46.53 ±6.05) points and (36.35 ±5.18) points,respectively,which were lower than those in the control group [(83.1 ±6.58)points,(81.85 ±6.70)points and (76.86 ±76.86)points](t＝31.369,29.279,38.464,all P＜0.05).After treatment for 1 week,2 weeks,the classification scores in the observation group were (4.21 ±2.03) points and (5.35 ±2.23) points,respectively,which were significantly higher than those in the control group [(3.35 ±1.24)points and (3.95 ±1.24)points] (t＝2.705,4.106,all P＜0.05).The effective rate was 91.0%in the observation group ,which was 89.2%in the control group,there was no statistically significant difference (χ2＝0.022,P＞0.05).Conclusion Olanzapine combined with rTMS is effective in the treatment of schizophrenia ,and olanzapine combined with rTMS is more effective in improving the cognitive ability of patients than olanzapine alone .

BACKGROUNDIt has been known that there are declines of dendritic cell (DC) count and its function in the peripheral blood of patients with non-small cell lung cancer (NSCLC), which are remarkably related to the proceeding and prognosis of NSCIC. The aim of this study is to investigate the relationship and clinical significance between the DC subsets (CD11c+DC and CD123+DC) and immunology state of patients with advanced NSCLC.

METHODSFlow cytometry was used to detect DC subsets, T cell subsets, NK cell percentage in the peripheral blood of 40 patients with advanced NSCLC and 10 healthy controls.

RESULTSThe expression of CD11c+DC (0.38%±0.18%) in the peripheral blood of advanced NSCLC patients, was decreased significantly compared with that of normal controls (0.66%±0.24%) (P < 0.01). The expression of CD123+DC in the peripheral blood of advanced NSCLC patients was (0.28±0.17)%, with no significant difference compared with that of normal controls (0.27%±0.11%) (P > 0.05). The percentage of CD3+ T cell and CD4+/CD8+ of patients with advanced NSCLC were significantly lower than those of normal controls (P < 0.01 and P < 0.05), and the percentage of CD8+ and NK cell of patients were higher than those of normal controls (P < 0.05). The correlation analysis showed that CD123+DC percentage was positively correlated to CD3+T cell percentage (P < 0.05) and negatively correlated to NK cell percentage (P < 0.01). The expression of DC subsets correlated with KPS and chemotherapy history of patients (P < 0.01).

CONCLUSIONSThe advanced NSCLC may induce significant decreasing expression of CD11c+DC and may induce significant decrease of immunology state. There are close relationship among the expression of DC subsets and the immunology state of patients as well as the clinical biological behaviors of patients with advanced NSCLC.

Objective To investigate the correlation between thyroid function,neutrophil/lymphocyte ratio(NLR),platelet/lymphocyte ratio(PLR)and diabetic ketoacidosis(DKA).Methods A total of 272 diabetic patients admitted to the Department of Endocrinology,Anhui No.2 Provincial People's Hospital were enrolled in this study from January 2021 to March 2024 and divided into DKA group(n=74),diabetic ketosis group(DK group,n=100)and simple DM group(n=98)according to their disease conditions.Serum free triiodothyronine(FT3),free tetraiodothyronine(FT4),thyroid stimulating hormone(TSH),FPG,HbA1c,anion gap(AG),white blood cells(WBC),neutrophils(N),platelets(PLT)and lymphocytes(L)were detected in each group,and NLR and PLR were calculated.The influence factors of DKA were analyzed by logistic regression,and the diagnostic value of FT3,NLR and PLR in DKA was evaluated by receiver operating characteristic(ROC)curve.Results Hospitalization days,FPG,HbA1c,incidence of LT3S,AG,WBC,N,PLT,NLR,PLR were higher in DKA group than in DM and DK groups(P<0.05),Age,FT3,FT4,TSH,L were lower in DKA group than in DM and DK groups(P<0.05).Logistic regression analysis showed that FT3,AG,NLR and PLR were the influencing factors for DKA.ROC curve showed that the area under the curve of FT3,NLR,PLR and combined detection to predict DKA were 0.961,0.938,0.810 and 0.980,the sensitivity was 87.8%,90.5%,75.7%and 98.6%,the specificity was 95.5%,92.9%,76.3%,92.4%,and the combined detection was superior to the single detection.Conclusions FT3,NLR,PLR have a certain diagnostic value for DKA,and the combined detection of the three has a high diagnostic value,which can provide a reference for clinical diagnosis of DKA.

Objective:To explore the effects of Xuebijing injection and its component hydroxysafflor yellow A on coagulation and survival rates of septic rats.Methods:① Assessment of coagulation: 144 male Sprague-Dawley (SD) rats were divided into four groups by random number table: sham group, cecal ligation and puncture (CLP) induced sepsis model group (CLP group), CLP+Xuebijing group, and CLP+hydroxysafflor yellow A group, with 36 rats in each group. CLP was used for reproducing septic models. The cecum of the rats in the sham group was exposed by laparotomy and then returned to the abdominal cavity without CLP, while the other steps were the same as those in the CLP group. Rats in the CLP+Xuebijing group and CLP+hydroxysafflor yellow A group were injected with Xuebijing (4 mL/kg, twice a day) or hydroxysafflor yellow A solution (0.378 g/L, 298 μg each time, twice a day) through caudal vein after operation. Levels of prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), fibrinogen (Fib), and D-dimer in peripheral blood were measured by automatic coagulation analyzer at 6, 12, 24 hours after operation. The enzyme linked immunosorbent assay (ELISA) was applied to determine levels of tissue factor (TF), tissue factor pathway inhibitor (TFPI), and soluble thrombomodulin (sTM) in peripheral blood. ② Analysis of survival rates: 120 rats were divided into four groups by random number table (the same groups with those in the section of assessment of coagulation), with 30 rats in each group. The Kaplan-Meier survival curve was plotted, and the cumulative survival rates were observed and recorded for 7 days after CLP surgery.Results:① Results of coagulation assessment: compared with the sham group, septic rats in the CLP group showed significant dysfunction in coagulation early, as evidenced by prolonged PT at 6 hours after CLP (s: 8.9±0.2 vs. 8.4±0.4, P < 0.01), and significantly increased levels of Fib, D-dimer, TFPI and sTM [Fib (g/L): 2.8±0.3 vs. 2.3±0.1, D-dimer (ng/L): 1.8±0.2 vs. 1.5±0.1, TFPI (ng/L): 131.1±10.9 vs. 102.8±10.5, sTM (μg/L): 27.2±1.2 vs. 19.8±2.9, all P < 0.01]. The coagulation dysfunction became more and more serious at 12 hours after operation, and further deteriorated with time. The use of both Xuebijing and hydroxysafflor yellow A revealed significant improvement in coagulation of septic rats at 6 hours, as shown by shortened PT (s: 8.3±0.2, 8.3±0.1 vs. 8.9±0.2, both P < 0.01), and decreased Fib, D-dimer, TFPI and sTM as compared with those in the CLP group [Fib (g/L): 2.3±0.1, 2.3±0.2 vs. 2.8±0.3; D-dimer (ng/L): 1.5±0.1, 1.5±0.2 vs. 1.8±0.2; TFPI (ng/L): 109.5±10.2, 91.5±5.0 vs. 131.1±10.9; sTM (μg/L): 22.3±1.5, 21.1±1.8 vs. 27.2±1.2; all P < 0.01]. However, there was no significant difference in coagulation function between the two intervention groups. ② Results of survival rates analysis: the rats in the sham group all survived 7 days after operation. The 7-day cumulative survival rate of the CLP group was only 36.67% (11/30). Compared with the CLP group, the cumulative survival rates were significantly increased in rats of the CLP+Xuebijing group and CLP+hydroxysafflor yellow A group [66.67% (20/30), 66.67% (20/30) vs. 36.67% (11/30), both P < 0.05], but no significant difference was found between the CLP+Xuebijing group and CLP+hydroxysafflor yellow A group. Conclusion:Both Xuebijing and its component hydroxysafflor yellow A appear to be capable of alleviating coagulation disorders and improving survival rates of septic rats effectively, and the effects show no significant difference between them.