Objective To study the mucosal and systemic immune response after intranasal immunization with mucosal complex vaccine for Toxoplasma gondii,and to observe the protective effect on mice. Methods The mucosal complex vaccine was made of soluble tachyzoite antigen (STAg) and cholera toxin (CT),which were mixed and dissolved in PBS (1 ml PBS containing 1 mg STAg and 50 ?g CT). Fifty-two BALB/c mice were randomly divided into two groups: immunized group and control. Mice were intranasally immunized with 20 ?l mucosal complex vaccine (20 ?g STAg and 1?g CT) per mouse twice at an interval of two weeks,while the control mice were given PBS solution instead. Six mice of each group were killed by dislocation of cervical vertebra on day 14 after the last immunization. The specific IgG antibodies in serum and IgA in feces were detected by ELISA. Lymphocytes in spleen,Peyer's patches (PP) and intestinal intraepithelial lymphocyte(IEL) were isolated and counted. Percentage of CD4+ and CD8+ T cells was determined by immunocytochemistry. Other mice were challenged intragastrically each with 4?104 tachyzoites of RH strain Toxoplasma gondii on day 14 after the last immunization. Their health condition was observed and the number of tachyzoites in liver and brain was determined microscopically on the 30 th day after challenge. Results IgG antibodies in serum and IgA antibodies in feces of immunized mice were higher than the control (P

Objective To compare the measured resting energy expenditure (MREE) in patients with sepsis calculated using respiratory indirect calorimetry and Harris-Benedict (HB) coefficient method,and to investigate the influence of different energy target on the prognosis of patients with sepsis.Methods This was a prospective comparative study.60 patients with sepsis who were suitable for nutrition support and respiratory indirect calorimetry in the Intensive Care Unit (ICU) of the General Hospital of Ningxia Medical University from January to October 2015 were selected.(1) MREE was measured simultaneously with respiratory indirect calorimetry and HB coefficient (Harris-Benedict equation × stress coefficient) in the 60 patients on the 0,3rd,7th,and 14th days of nutrition support.The MREE calculated with the two methods were compared to detect difference.(2) The septic patients were randomly divided into two groups according to the random number table:respiratory indirect calorimetry group (n =30),in which the nutrition support was given according to the MREE measured using respiratory indirect calorimetry as the energy target;HB coefficient method group (n =30),in which the nutrition support energy target was the MREE measured using HB coefficient method.The clinical data of the patients in both groups were recorded and compared,which included the baseline characteristics,nutrition-related complications,and prognostic indicators.Results (1) Within 2 weeks during nutrition support,the dynamic MREE in the respiratory indirect calorimetry group was significantly higher than that in the HB coefficient method group (P ＜0.05).There was no significant difference in MREE on the 0,3rd,7th,and 14th days of nutrition support measured by respiratory indirect calorimetry (P ＞ 0.05).(2) There were no significant differences between the two groups in incidence of liver function damage and abnormal blood glucose within 2 weeks of nutrition support (both P ＞ 0.05).Compared with the HB coefficient method group,the length of ICU stay and 28-day mortality were significantly lower in the respiratory indirect calorimetry group [193.5 (172.2,289.7) h vs.247.5 (194.7,393.2),Z=-2.061,P=0.039;3.3％ vs.23.3％,x2 =5.129,P=0.023],but there was no significant difference between the two groups in mechanical ventilation time (P ＞ 0.05).Conclusions The MREE of septic patients may be considerably high and with little fluctuation in a short period.Respiratory indirect calorimetry may be more suitable to guide the energy intake target in septic patients.

Objective To observe dynamically the location and time of attachment and invasion of Toxoplasma gondii tachyzoites to murine intestinal mucosa by chromogenic in situ hybridization targeting SAG2 mRNA. Methods Thirty 7- to 8-week-old BALB/c mice were randomly divided into experiment group(24 mice)and control group(6 mice). Each animal in the experiment group was given 2?104 tachyzoites of RH stain in 0.2 ml PBS by intragastric administration and that in the control group was given 0.2 ml PBS. Four mice in the experiment group and one in the control group were sacrificed at 15 min,30 min,1 h,2 h,4 h and 8 h after infection,respectively,and paraffin sections of duodenum,jejunum and ileum were prepared to perform the in situ hybridization with Dig-labeled oligonucleotide probe complementary to SAG2 mRNA of T. gondii. Results Tachyzoites were found on the striated border of small intestine epithelial cells (absorptive cells,goblet cells and endocrine cells),in or between two absorptive cells or in the lamina propria. At 15 min-2 h after infection,there was significant difference in the number of attachment on jejunum and ileum (P

To develop a low-cost, reliable, easy-to-maintain and practical instrument for protein purification. Some ultraviolet luminescent diode was used to provide 280 nm light source, and high-sensitivity S1336 photo-electric detector was employed for real-time monitoring of purified protein solution flowing through quartz cell to supervise the concentration of the protein. The instrument gave voice alarm and stopped working in case the protein concentration was less than the standard one. The lower SCM monitored the liquid level of the protein collecting cup and the position of loading arm through laser infrared distance sensor, so that a cup full of protein might be replaced by another empty cup. The instrument involved in Samsung S5PV210 embedded master computer, Wince6.0 operating system, Keil4.0 and VS2005. Trials proved that the instrument could perform real-time monitoring and curve display of dual-channel ultraviolet absorption, and could realize auto collection of 735 ml protein solution and up to 5-hour standby. The instrument developed has simple structure, high reliability and easy maintenance, and meets the desired require-ments.

Objective To investigate the clinical significance of Notch-1 expression in small cell lung cancer(SCLC) and non-small cell lung cancer(NSCLC),and analyze the role of it in prognosis.Methods SP immunohistochemistry was used to detect the expression of Notch-1 antibody in 43 of SCLC and 40 of NSCLC tissues.Further analysis was carried out to interpret the association of Notch-1 antibody expression with clinicopathological features,lymph node metastasis and prognosis in SCLC.Results The positive rate of Notch-1 expression was 20.93％ ( 9/43 ) in SCLC,while 65.00％ ( 26/40 ) in NSCLC.The expression of Notch-1 antibody was associated with clinical stage and lymph node metastasis ( x2 =5.42,P ＜ 0.05 ; x2 =4.88,P ＜ 0.05respectively),but was not associated with age,sex,tumor location,tumor size ( Ps ＞ 0.05 ).Compared with NSCLC,the expression rates of Notch-1 antibody were significantly higher in SCLC ( x2 =16.50,P ＜ 0.05 ).Kaplan-Meier survival analysis indicated that the survival time of patients with positive Notch-1 expression was significantly longer than that of patients with negative staining( x2 =19.87,P ＜ 0.05 ).Cox regression analysis showed that Notch-1 antibody could significantly reduce the risk of death in patients with SCLC.Conclusion The positive expressions of Notch-1 were significantly different in SCLC and NSCLC,which linked to the clinicalstage,lymph node metastasis and poor prognosis.Accordingly,the expression of Notch-1 may have good value in diagnosis and prognosis.

AIM:To explore the polarized distribution of M 2 macrophages in the marginal region around lung adenocarcinoma , the marginal/central ratio and their effect on the prognosis .METHODS:Double immunohistochemistry staining was used to determine the distribution and the difference of CD 163 +/CD68 +( M2 ) macrophages in the marginal and central regions in 49 cases of lung adenocarcinoma in situ ( AIS), 11 cases of minimally invasive adenocarcinoma ( MIA) and 57 cases of invasive adenocarcinoma ( IA) in order to explore the effect and mechanism of the polarized distri-bution and the marginal/central ratio on the progression of lung adenocarcinoma .Single-factor Kaplan-Meier survival curve analysis and multivariate Cox survival analysis were employed to explore the relationship between the polarized distribution of M2 macrophages and the prognosis .RESULTS:Polarized aggregation of M 2 macrophages was observed in the marginal region of lung adenocarcinoma compared with that in the central region , and the difference was significant ( P<0.01 ) . Based on the median level , they were divided into high polarized group and low polarized group .In low polarized group , M2 macrophage count in AIS was not significantly different from that in MIA or IA .However, in high polarized group, M2 macrophage count in AIS was lower than that in MIA and IA in turn and there were statistically significant differences (P<0.01).Single-factor Kaplan-Meier survival curve analysis and log-rank test result showed that the number of M2 mac-rophages in the marginal region and marginal/central ratio were negatively correlated to the survival time (χ2 =44.71, P<0.01;χ2=21.75, P<0.01).Multivariate Cox survival analysis showed that the high polarized distribution of M 2 macro-phages in the marginal region and the marginal /central ratio were independent risk factors for the prognosis ( P<0.01 ) . CONCLUSION:There is a polarization effect of M2 macrophages on the marginal region of lung adenocarcinoma .The marginal polarization and the marginal/central ratio are independent risk factors of the prognosis .Therefore , it may be an effective method for the evaluation of the prognosis to judge the marginal polarization by preoperative puncture and to deter -mine the marginal/central ratio of M2 macrophages by postoperative biopsy .

Objective: To observe the effect of reinforcing and reducing manipulation by breathing on yin and yang meridian points for motor function of knee joints in patients with ischemic stroke. ＜br> Methods: A total of 60 ischemic stroke cases were randomly allocated into an observation group and a control group, 30 in each group. Other than basic and conventional rehabilitation therapies, cases in the observation group were treated with reinforcing and reducing manipulation on yin and yang meridians points for a period of 8 weeks; whereas cases in the control group only received the same basic and conventional rehabilitation therapy as that in the observation group. Then the knee joint energy, support moment and shaking moment were compared between the two groups before treatment, and after 4-week and 8-week treatment respectively. ＜br> Results: Before treatment, there were no between-group significant differences in knee joint energy, support moment and shaking moment (all P>0.05). After treatment, the knee joint energy, support moment and shaking moment in both groups were significantly improved (P＜0.05). After 8 weeks of treatment, the knee joint energy, support moment and shaking moment in both groups were significantly improved than that of 4-week treatment (P＜0.05). In addition, there were between-group statistical significances in comparing knee joint energy, support moment and shaking moment after 4 and 8 weeks of treatment (P＜0.05). ＜br> Conclusion: Reinforcing and reducing manipulation by breathing on yin and yang meridian points can increase the knee joint support and shaking force and moment in stroke patients and improve their walking ability.

Objective To observe the change of extension-flexion arc of ankle in hemiplegia due to ischemic stroke after yin-yang acupuncture associated with respiration needling manipulations.Method Sixty ischemic stroke patients were divided into a control group and a treatment group by random number table, 30 in each group. The control group was intervened by basic treatment, while the treatment group was by yin-yang acupuncture associated with respiration needling manipulations in addition to the basic treatment. Before intervention and 28 d after intervention, the Lokomat gait analysis system was adopted to observe the change of the extension-flexion arc of ankle respectively under motorial and stationary states.Result When heel was grounded, the flexion angle, the maximum extension-flexion arc value, and motility of ankle joint of the treatment group were statistically different from those of the control group (P＜0.05).Conclusion Yin-yang acupuncture associated with respiration needling manipulations can effectively improve the movement and the walk function of hemiplegia due to ischemic stroke.

Objective To observe the effect of yin-yang acupuncture in association with respiratory reducing-reinforcing needling manipulations on the gait cycle in patients with post-stroke hemiplegia. Methods Sixty patients with ischemic stroke were randomized into a treatment group and a control group, 30 each. The control group was intervened by conventional medication, while based on which the treatment group was additionally given yin-yang acupuncture in association with respiratory reducing-reinforcing needling manipulations. The kinematic time parameters were evaluated before intervention and after 28-day treatment. Results After intervention, the stance phase of the affected side and the healthy side, ambulation cycle, swinging phase of the affected side and the healthy side, and the stance phase of both limbs in the treatment group were significantly different from that in the control group(P＜0.05). Conclusion Yin-yang acupuncture in association with respiration reducing-reinforcing needling manipulations can effectively improve the gait cycle and ambulation function of the patients with post-stroke hemiplegia.