Objective To construct the prokaryotic expression vector of mouse CD25 extracellular domain and to express it in E coli.Methods Total RNA was isolated from splenocytes of Balb/c mice.The CD25 extracellular domain gene was amplified by RT-PCR and cloned into the PET-32a vector.A positive recombinant,PET-32a-CD25e,was identified by enzyme cleaving and sequencing before its expression in E.coli,and transferred into E.coli BL21(DE3) plysS for its expression.After purification with Ni+ resin and renaturation in vitro,a relative molecular mass(Mr) of the interesting protein was detected by SDS-PAGE and Wes-tern blotting.Effect of the purified interesting protein on the proliferation of splenocytes from T cell vaccine-immunized syngeneic mice was detected by MTT assay.Results The cloned CD25 extracellular domain gene was identified to be functional by sequencing and expression.The purified interesting protein could significantly induce the proliferation and IL-4 secretion of splenocytes from T cell vaccine-immunized mice in vitro.Conclusion Mouse CD25 extracellular domain gene can be successfully expressed in prokaryotic cells with biological activity,which lays a foundation for further relative studies.

The structure, reaction mechanism and molecular biology of Leuconostoc mesnteroides dextransucrase are reviewed.

Objective To monitor the quality changes of iodized salt and analyze its impact factor in Chongqing between 2001 and 2009. Methods Salt samples were collected according to the east, west, south,north and center locations in iodized salt production, wholesale and household sectors. Two units in iodized salt production and wholesale segment were sampled from north, south, east and west places and only 1 unit was sampled from the central place. Nine samples were collected every month in each place. If the place had less than 9 units, and then taken all the units. About resident household, 2 townships were sampled from north, south, east and west places, and 1 township was sampled from the central place, then 20 samples were collected from each township. Iodine content was detected by oxidation-reduction assay. The index of mean iodine, qualified rate from factories and wholesale, coverage rate and taking rate of qualified iodized salt in residents were calculated.Significance was analyzed by trend test, analysis of variance and X2 test. Results The qualified rate of iodized salt from the manufacturers was 92.9%(13/14) in 2001 and the rate was 100.0% each year from 2002 to 2009. The qualified rates of iodized salt from the wholesale were 88.7%(282/318) - 99.8%(431/432). The rates of 2001 and 2002 were lower than that of other years(X2 = 4.98 - 45.69, all P< 0.05 or < 0.01). The coverage rate and taking rate of qualified iodized salt in residents were 94.2% (11 154/11 841 ) - 98.9% ( 14 061/14 217), 83.5% (9 887/11 841 ) -95.8% (13 449/14 039), respectively. The rates showed an increasing tendency (F = 9.27, 26.39, all P < 0.05).The districts(counties) with qualified iodized salt consumption rate > 90% kept increasing. The mean iodine from the manufacturers and wholesale were 29.71 - 36.25, and 31.26 - 36.13 mg/kg, respectively. The iodine level showed a descending trend(F = 35.45, 140.59, all P < 0.01 ). The mean iodine level from the inhabitants were 28.84 - 30.98 mg/kg which remained stable (F = 3.05, P > 0.05 ). The iodine level from manufacturers, wholesale to inhabitants showed an descending trend(F = 38.46 - 671.23, all P < 0.01 ). Conclusions The surveillance results of iodized salt shows an increasing tendency in quality of iodized salt, eoverage rate and taking rate of qualified iodized salt. Factors that affect the quality of iodized salt is that the enterprise does not add iodine to salt strictly by the standard.

Objective:To observe the effect of warm-unblocking acupuncture plus fluticasone propionate nasal spray on the pulmonary ventilation, level of interferon-γ (IFN-γ) and sleep quality in patients with allergic rhinitis (AR). Methods: A total of 112 AR patients were enrolled between January 2013 and August 2018 and were divided into an observation group and a control group by the random number table method, with 56 cases in each group. Patients in the observation group received warm-unblocking acupuncture plus fluticasone propionate nasal spray, and patients in the control group only received fluticasone propionate nasal spray. The nasal symptom score, pulmonary function indexes, the levels of IFN-γ and interleukin (IL)-4 in serum, and sleep quality in the two groups were compared. Results: After treatment, the total effective rate in the observation group was higher than that in the control group (P＜0.05). The nasal symptom score dropped in both groups after treatment (both P＜0.05), and the score in the observation group was lower than that in the control group (P＜0.05). The pulmonary ventilation indexes all increased significantly after treatment in the observation group (all P＜0.05); the forced expiratory volume in 1 second (FEV1) to forced vital capacity (FVC) ratio (FEV1/FVC) and the forced expiratory flow at 50%, 75% and 25%-75% of the vital capacity (FEF50%, FEF75%, FEF25%-75%) increased after treatment in the control group (all P＜0.05); the pulmonary ventilation indexes were higher in the observation group than those in the control group (all P＜0.05). The level of IFN-γ increased significantly after treatment in the two groups (both P＜0.05) and the level of IL-4 dropped significantly (both P＜0.05); the observation group had a higher IFN-γ level (P＜0.05) and a lower IL-4 level (P＜0.05) compared with the control group. Regarding the Pittsburgh sleep quality index (PSQI), the scores of subjective sleep quality, habitual sleep efficiency and sleep disturbances and the general PSQI score decreased significantly after treatment in both groups (all P＜0.05), and the scores in the observation group were significantly lower than those in the control group (all P＜0.05). Conclusion: Warm-unblocking acupuncture plus fluticasone propionate nasal spray can effectively control the clinical symptoms and improve pulmonary function in the treatment of AR; this approach can regulate the levels of IFN-γ and IL-4 towards the normal range in AR patients; it can also improve patient’s sleep quality. This method can produce more significant efficacy than fluticasone propionate nasal spray used alone.

Objective To analyze the characteristics of urinary iodine and edible iodized salt,and to provide suitable iodine concentration in iodized salt.Methods Sample testing was carried out to detect iodine concentration in marketed salt and child urine before salt iodization (1994).After salt iodization (2001-2010),sample testing was carried out to detect salt iodine level in manufacture,market(2001-2010) and resident household(1997-2010).Urine of children aged 8 to 10 were sampled by PPS method from 1997 to 2005.In 2009,urinary iodine of 20 children was determined in each of 5 schools,which were sampled from 5 counties located at North,South,East,West and center of Chongqing.Based on the urinary iodine and salt iodine levels before salt iodization,the relationship of urinary iodine and consumption of iodized salt was calculated.Suitable iodine concentration in iodized salt was put forward.x2 test and trend analyze approach(F-test) were used for statistical analysis.Results Before salt iodization,salt iodine level was not tested in 204 edible salt samples; the median of urinary iodine was 53.14 μg/L in 1374 children.After salt iodization,form 2001 to 2010,the average iodine levels in manufacture and wholesale salt were between 29.72-36.25 mg/kg and 30.65-36.13 mg/kg,respectively,both of them decreased significantly(F =35.35,140.59,all P ＜ 0.01),and show a downward trend.Batch quality passing rate of industry iodized salt was 100％ except in 2001,which was 92.86％.Batch quality passing rate of market iodized salt were between 88.68％-99.77％,specifically in 2001 (88.68％),in 2002(92.57％) and in 2003 (96.22％).There was no significant difference in other years (all P ＞ 0.05).The median of urinary iodine were between 238.80-328.00 μg/L,more than 35％ fall into ＞ 300 μg/L; while salt iodine increased 1 mg/kg,urinary iodine increased 5.51 μg/L-7.40 μg/L; The medium of urinary iodine of children were between 140.05-383.00 μg/L in 40 counties or districts in 2009.Reducing the iodine concentration in edible iodized salt to 20 mg/kg,the median of urinary iodine can be kept at 163.34 μg/L to 201.14 μg/L.Conclusions Iodine in iodized salt is above sufficient in Chongqing.Salt iodine should be reduced to 20 mg/kg,which will meet various population's need.

Objective To find out the current thyroid volume of school children and its influencing factors in Chongqing.Methods Probability sampling method(PPS) was used to select 30 counties in Chongqing in 2011.Forty children aged 8-10of 1 randomly selected school from every county chosen were enrolled in the study.Thyroid volume of children was examined by B-ultrasonograghy.Body height and body weight were measured.The relationship between gender,age,height and weight and thyroid volume was analyzed,respectively.Results One thousand three hundred and twenty-two children aged 8-10 were investigated.The median of thyroid volume was 3.53 ml.The goiter rate was 5.52％ (73/1322).Thyroid volume of female and male was 3.55 and 3.51 ml,respectively.There was no significant difference of thyroid volume between female and male (H =0.68,P ＞ 0.05).Thyroid volume of children aged 8,9 and 10 was 3.30,3.53 and 3.76 ml,respectively.There was a significant difference of thyroid volume among different age groups(H =52.49,P ＜ 0.01).Thyroid volume of children height (110-,120-,130-and ≥140 cm,respectively) was 2.96,3.22,3.59 and 4.13 ml.There was a significant difference of thyroid volume among different height groups (H =149.23,P ＜ 0.01).Thyroid volume of children weight(17-,20-,30-and ≥40 kg,respectively) was 2.71,3.31,3.91 and 4.74 ml.There was a significant difference of thyroid volume among different weight groups(H =138.44,P ＜ 0.01).For the coefficients of simple and partial correlation,there was a significant correlation between thyroid volume and age,height and weight (P ＜ 0.05).The Spearman coefficient was 0.2411,0.3950 and 0.4285,respectively.The partial correlation coefficient was 0.0640,0.1154 and 0.2319,respectively.The standard partial coefficient of age,height and weight was 0.640,0.1154 and 0.3410,respectively.The proportion of the standard partial coefficients was 1 ∶ 1.8 ∶ 5.3.The function of body weight to thyroid volume was 5.3 times that of age and 3.0 times that of body height.Conclusions The goiter rate of schoolchildren in Chongqing is relatively high.Thyroid volume is affected by age,body height and body weight.The relationship between thyroid volume and iodine nutrition needs further study.

Objective To investigate the nutritional status of iodine among residents in Chongqing, and to facilitate scientific prevention and control of iodine deficiency disorders. Methods Select 9 towns in each of the 40 districts (counties) in Chongqing, and collect 40 resident edible salt samples in each of the selected town to detect salt iodine by direct titrimetry. Select 5 towns on the site of the east, west, south, north and middle of every district (county), select 20 children aged 8 to 10 in each of the selected town to collect urine samples and detect urinary iodine by As-Ce catalytic spectrophotometric assay. Results The median of iodine of 14 217 salt specimens by household was 292 mg/kg with a coverage rate of qualified iodized salt of 98.90%( 14 061/14 217). The consumption rate of qualified iodized salt was 95.59%( 13 590/14 217). The median of urinary iodine for 4050 children aged 8 to 10 was 247.20μg/L, of which < 50 μg/L accounted for 4.60%(186/4050), 50-99μg/L accounted for 7.28% (295/4050), 100 - 199 μg/L accounted for 26.44% (1071/4050), 200 - 299 μg/L accounted for 25.58% (1036/4050), 300 μg/L or more, accounted for 36.10% (1462/4050). However, no significant difference was observed between different age groups(x2 = 3.77, P > 0.05). At district (county) level, the median of urinary iodine in 10(25.00%) districts (counties) was 100 - 200 μg/L, that in other 23(57.50%) districts (counties) was 200 - 300 μg/L, and that in other 7(17.50%) districts/counties was greater than 300 μg/L, and statistical significance was observed between different districts/counties (x2 = 441.95, P < 0.01). Conclusions Current iodine nutrition among residents in Chongqing is adequate. While there is excess, need to reduce the amount of salt iodization.

Objective To analyze the spatial distribution characteristics of non-iodized salt at household level based on geographic information system (GIS) in Chongqing.Methods The database of non-iodized salt at county level from 2001 to 2010 was established in Chongqing.By using GIS technology,the spatial distribution and spatial autocorrelation were analyzed by ArcGIS 9.3 software.Results The rate of non-iodized salt was fluctuated between 2.35％-5.78％ during 2001-2006 and the rate was reduced to less than 2.00％ after 2007.The result of spatial autocorrelation analysis on non-iodized salt from 2001 to 2006 indicated that Moran's Ⅰindex was 0.145578,0.078801,0.108033,0.091957,0.127749,0.214302,respectively(Z value was 3.066275,1.977321,2.541619,2.309972,2.900446,3.874203,respectively,all P ＜ 0.05).The spatial distribution of non-iodized salt had marked spatial cluster through Chongqing region from 2001 to 2006.The result of local spatial autocorrelation analysis from 2001 to 2006 indicated that Fengdu and Fuling were two high-risk areas(all P ＜ 0.05).Dianjiang,Yubei,Jiangbei,Wulong and Banan were also confirmed as high-risk areas in 2001,2005 and 2006(all P ＜ 0.05).The results also indicated that the distribution of non-iodized salt in the seven high-risk areas was positively correlated.The result of spatial autocorrelation analysis on non-iodized salt from 2007 to 2010 indicated that Moran's Ⅰ index was 0.018361,0.016186,0.040769,-0.059691,respectively (Z value was 1.093310,0.787361,1.071811,-0.583820,respectively,all P ＞ 0.05).The spatial distribution of non-iodized salt was at random on the whole from 2007 to 2010.However,there were four local high-risk areas.The distribution in Fengdu and Dianjiang was positively correlated,while that in Jiangjin and Shizhu was negatively correlated.Conclusions The distribution of non-iodized salt at households level in Chongqing is changed from spatial distribution before 2006 to random distribution after 2007,but there are high value areas,which should be taken as the focus of monitoring.

OBJECTIVETo investigate the specific antitumor immune response induced by idiotype protein (Id)-pulsed dendritic cells (DC) in vitro.

METHODSDC was generated from peripheral blood monocytes of the multiple myeloma (MM) patients using GM-CSF, IL-4, and TNF-alpha. The DCs were pulsed with idiotypic fragment, the F(ab')2 fragment of M protein from MM patient at the immature stage. The morphologic characteristics of the cells were observed with light and electron microscopes. The phenotypic features were analyzed with FACS, MTT assay was employed to evaluate the proliferation of autologous T cells and the inhibition rate of MM cells.

RESULTSDC precursors in peripheral blood could be induced to typical mature DC in medium containing GM-CSF, IL-4 and TNF-alpha. Mature DC with Id could increase the proliferation of the autologous T cells and activate naive T cells to become tumor specialized cytotoxic T lymphocytes (CTL). The CTL at different doses showed significant inhibition on or killing ability to autologous MM cells in vitro.

CONCLUSIONSIn a suitable cytokine environment, the DC precursors from peripheral blood of MM patients could be induced to functional DC, and vaccination of Id-pulsed DC could induce active antitumor immune response.

Adult ; Aged ; Antibodies, Anti-Idiotypic ; immunology ; Cancer Vaccines ; immunology ; Cells, Cultured ; Dendritic Cells ; immunology ; Female ; Humans ; Immunotherapy, Active ; Male ; Middle Aged ; Multiple Myeloma ; immunology ; therapy ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo study the effects of intestinal microflora alteration on specific and nonspecific immune function and hematopoietic function of mice.

METHODSSixty BALB/C mice were divided at random into two groups, experimental group and control group, with 30 mice in each. The mice in the experimental group were given kanamycin 50 mg while those in the control group were given distilled water intragastrically everyday for consecutive 10 days. After the 10 day treatment all the mice were sacrificed, and the cecal contents were collected for quantitative analysis of the intestinal bacterial flora. Certain indexes of immune function, including phagocytosis rate of macrophages, number of T lymphocytes positively stained by esterase and serum interleukin 2 (IL-2) content, and the weight of the spleen, granulocyte-macrophage colony stimulating factor etc. as indexes of hematopoietic function were determined.

RESULTSIn the group, the quantity of Enterobacteriaceae, Enterococcus, Bifidobacterium and Lactobacillus were significantly lower than that in the control group (P < 0.01). The number of PFC (plaque forming cells), the phagocytosis rate of macrophage, the number of T lymphocytes with positive NANE staining, the level of IL-2 significantly decreased when compared with that in the control group (P < 0.01). The weight of the spleen in the experimental group decreased when compared with that in the control group (P < 0.01). Levels of IL-3, GM-CSF, the total number of WBC and the proportion of neutrophil remarkably decreased as compared to that in the control group (P < 0.01). Analysis of the correlations between normal microflora, immunologic and hematopoietic indexes showed that marked positive correlations between the quantity of Bifidobacteria and each immune index including the levels of IL-3 and GM-CSF. There was a positive correlation between IL-2 and IL-3, IL-2 and GM-CSF as well.

CONCLUSIONThe application of antibiotics may cause changes in the structure and quantity of intestinal microflora. The dysbacteriosis may decrease the immune function of organism. The dysbacteriosis may decrease the hemopoietic function. The dysbacteriosis, the decrease in immune and hematopoietic function may affect one another. The balance in microecosystem should be emphasized and antibiotics should be applied rationally to reduce the side effects such as dysbacteriosis.

Animals ; Anti-Bacterial Agents ; pharmacology ; Esterases ; biosynthesis ; Feces ; microbiology ; Granulocyte-Macrophage Colony-Stimulating Factor ; analysis ; Interleukin-2 ; blood ; Intestines ; drug effects ; microbiology ; Kanamycin ; pharmacology ; Macrophages ; drug effects ; physiology ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Organ Size ; Phagocytosis ; drug effects ; Spleen ; drug effects ; pathology ; T-Lymphocytes ; drug effects ; metabolism