Objective To determine the immune responses induced by recombinant Salmonella ty-phimurium expressing the secreting antigen ESAT-6 of Mycobacterium tuberculosis. Methods ESAT-6 cod-ing gene was cloned and identified by PCR and sequencing. Prokaryotic expression plasmid pYA33-esat car-rying the ESAT-6 coding sequence was constructed firstly and electro-transformed into an attenuated strain X4550 of Salmonella typhimurium, the recombinant bacteria was named as X4550(33-esat). C57BL/6 mice were immunized intranasally (I. N) with 108 CFU recombinant bacteria at day 0 and 18. Cells from spleen, lung, mesenteric lymph node (MLN) and Peyer's patch (PP) were collected from mice after second immu-nization, and the specific IFN-γ-secreting cells and IL-4-secreting cells were detected by ELISPOT assay u-sing ESAT-6 peptide as stimulus. Furthermore, CTL effects were in vivo evaluated by CFSE assay. Results The results showed that cellular immune responses specific for ESAT-6 could be detected by ELISPOT assay. In lung and PP cells, immune responses against ESAT-6 were biased toward Th1 type, the frequency of IFN-γ-secreting cells was much higher than that of IL-4-secreting cells. In splenocytes and MLN cells, the anti-gen specific immune responses acted as Thl and Th2 balance, the frequency of IFN-γ-secreting cells was close to that of IL-4-secreting cells. CFSE assay indicated that recombinant bacteria could induce the high level of CTL effects specific for ESAT-6 peptide. Conclusion These results suggested that recombinant Sal-monella typhimurium X4550(33-esat) not only can induce cellular immune responses, but also can elicit specific CTL responses after I. N immunization. It also provided the useful information for the control of infec-tious disease of tuberculosis.

The herbs used as the material for traditional Chinese medicine are mostly planted at mountain areas where there is appropriate weather condition. Due to the complex terrain condition in mountain area, the planting at such places is always associated with a scattered pattern, which inevitably bring difficulty to the work of accurate measurement for the planting area. Advanced research about monitoring the planting area for medicine-herb under complex environmental condition is so critical and meaningful for developing the medicine-herb planting to a large scale and an industry. Therefore, this research, taking the planting of Herpetospermum pedunculosum in Luding county as an example, utilizes RS&GIS: using the image data from TM and ETM datum as the remote sensing information source, integrating GPS data from quadrat survey on the spots, and analyzing on a GIS platform, to extract the planting area of H. pedunculosum in Luding county. The results indicate that: (1) the method and technology used to estimate the planting area of H. pedunculosum are feasible and can be applied to monitoring the planting area of medicine-herb at mountain area under complex environmental conditions: (2) the planting area of H. pedunculosum in Luding county is estimated to be 857. 246 acres by the methods utilized in this research, which is closed to the value from local statistical data with an error range less than 5%; (3) in consideration of those inevitable errorrs generating from the procedures such as remote sensing data acquisition, data preprocessing, data interpretation, and so on, and the fact that the classification of remote sensing data, which must be implemented by those above procedures, was used directly to extract the herb type, the accuracy of area estimation for the herb is significantly influenced by the accuracy of classification for herb type, and the estimated area is inevitably associated with some error.
China ; Cucurbitaceae ; chemistry ; growth & development ; Geographic Information Systems ; Medicine, Tibetan Traditional ; Remote Sensing Technology ; methods

Objective To summarize the clinical features of 22 probands diagnosed with congenital muscular dystrophy (CMD),and to provide genetic counseling and prenatal diagnosis for 23 fetuses of these pedigrees.Methods Data of 22 CMD patients who were treated in the Pediatric Department of Peking University First Hospital during October 2006 to March 2016 were analyzed.Informed written consents for participation in this study were obtained from the parents or guardians.Prenatal diagnosis was performed using DNA samples extracted from fetal villus cells of 12 cases at 11-13 gestational weeks and amniotic fluid of 11 cases at 18-22 gestational weeks.Direct DNA sequencing by polymerase chain reaction (PCR) and multiplex ligation-dependent probe amplification (MLPA) were used to detect CMD-related gene mutations.Linkage analysis of short tandem repeats (STRs) was used to identify maternal blood contamination and biological parents.Results Thirteen out of the 22 probands with CMD were diagnosed with congenital muscular dystrophy type 1 A (MDC1A),and all of them carried compound heterozygous mutations in LAMA2 gene.Prenatal diagnosis of 13 fetuses from these pedigrees found that four fetuses were wild-type,seven were heterozygotes and two carried the same mutations as their proband.Three probands with LMNA-related congenital muscular dystrophy (L-CMD) carried de novo mutations in LMNA gene.In these pedigrees,two fetuses were wild-type and one whose mother was mosaicism carried the same mutations as the proband.One proband with Ullrich congenital muscular dystrophy carried compound heterozygous mutations in COL6A2 gene and the fetus of the same pedigree was wild-type.Five probands were diagnosed with α-dystroglycanopathies.And among them,two cases of muscle-eye-brain disease (MEB) carried compound heterozygous mutations in POMGnT1 gene and the fetuses of the two peidgrees were heterozygotes;one case of congenital muscular dystrophy type 1C (MDC1C) had compound heterozygous mutations in FKRP gene and the fetus carried the same mutations;one patient diagnosed with POMGnT1-related congenital muscular dystrophy with mental retardation (CMD-MR) carried compound heterozygous mutations in POMGnT1 gene,and the fetus was positive for the same mutations;one proband with POMT1-related CMD-MR was positive for compound heterozygous mutations in POMT1 gene and the results of prenatal diagnosis for two fetuses of this pedigree showed that the first fetus had the same mutations as the proband,while the second was heterozygote.Conclusions No effective therapeutic method is available for CMD.Therefore,accurate genetic counseling and prenatal diagnosis are necessary to prevent CMD child from birth.

OBJECTIVETo study the relations between anthropometric parameters [body mass index (BMI), percent of body fat, waist circumference] and blood lipids of urban children in Beijing and to compare the ability of prediction effect of waist circumference, percent body fat and BMI on lipid profiles.

METHODS852 children aged 9-10 were recruited with multi-stage sampling. Serum lipids of children were measured using automatic biochemical instrument. Fisher's exact test, trend test and median regression were used to analyze the data from 847 subjects with valid information.

RESULTS(1)With the rise of the BMI, percent body fat and waist circumference, total triglycerides (TG), total cholesterol (TC) and low density lipoprotein choleasterol (LDL-C) showed increasing trends while high density lipoprotein choleasterol (HDL-C) had a decreasing trend(P < 0.01). (2)TG was mainly influenced by waist circumference (t = 6.86), sex (t = 3.96) and percent of body fat (t = 1.85); TC was influenced by percent of body fat (t = 5.31); LDL-C was influenced by waist circumference (t = 4.65), height (t = -2.79) and percent age of body fat (t = 2.77); HDL-C was influenced by waist circumference (t = -12.24), sex (1= -5.83).

CONCLUSIONWith the aggravation of children's fatness (BMI, WC and PBF), serum lipids would increase and WC seemed to be the best predictor for lipids among BMI, WC and PBF.

Body Composition ; Body Height ; Body Mass Index ; Child ; China ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Humans ; Triglycerides ; blood ; Urban Population ; Waist Circumference

OBJECTIVETo investigate the food water sources of adults in the four cities of China in summer.

METHODSA total of 64 adults aged 18 - 60 yrs from four cities (Beijing, Shanghai, Chengdu and Guangzhou)were selected using convenient sampling method. The food samples were collected by using duplicate portion method and the water content of food samples were determined by using the national standard. The information on amounts and types of daily drinking soup was recorded by subjects using a quantitative measurement.

RESULTSA total of 63 subjects had completed the investigation. The median of daily water from food of subjects was 1157 ml, while water from staple food was 480 ml, the median ratio of which accounting for 41.8% of water from food. Water from dishes was 427 ml, the median ratio of which accounting for 37.9% of water from food. Water from soups was 133 ml, the median ratio of which accounting for 13.8% of water from food. Water from snacks was 0 ml, the median ratio of which accounting for 6.4% of water from food. The medians of water from staple food (384, 388 and 435 ml, respectively) in Beijing, Shanghai and Guangzhou was significant lower than that of Chengdu subjects' (900 ml, χ(2) = 21.27, P = 0.000). But the median proportion of water from staple food of subjects in Beijing (47.7%) was significantly higher than that in Chengdu (43.2%), Shanghai (42.9%) and Guangzhou (33.9%) (χ(2) = 8.69, P = 0.034). The median of amount and proportion of water from soups of subjects in Guangzhou (267 ml, 24.4%) and Chengdu(278 ml, 15.7%) was significantly higher than that in Shanghai (133 ml, 9.0%) and Beijing (100 ml, 5.8%) (amount: χ(2) = 22.52, P = 0.000;proportion: χ(2) = 16.27, P = 0.001).

CONCLUSIONThe staple food and dishes are the main sources of daily food water.

Adolescent ; Adult ; China ; Drinking ; Female ; Food Analysis ; Humans ; Male ; Middle Aged ; Seasons ; Surveys and Questionnaires ; Urban Population ; Young Adult

OBJECTIVESTo investigate total drinking water intake of adults in the four cities of China in summer.

METHODSA total of 1483 adults aged 18 - 60 yrs from Beijing, Shanghai, Chengdu and Guangzhou were selected using multiple-stage random sampling method. The information of amounts and types of daily drinking water was recorded by subjects for seven consecutive days using a quantitative measurement. The amounts and types of daily drinking water among different cities and between men and women or urban and rural was analyzed.

RESULTSThe median of daily total drinking water of subjects was 1488 ml, with significant difference among the four cities (1579, 1793, 1150, 1467 ml in Beijing, Shanghai, Chengdu and Guangzhou city, respectively, χ(2) = 154.31, P = 0.000). The median of daily drinking water was significantly higher in men (1679 ml) than women (1370 ml) (Z = 8.34, P = 0.000), but no significant difference was found between urban (1514 ml) and rural (1466 ml) daily drinking water median (Z = -0.81, P = 0.420). The median of daily consumption of plain water, tea and beverages were 786, 109, 186 ml, respectively. Among four cities, the highest consumption of plain water was in subjects of Guangzhou (917 ml), while the highest tea consumption in Shanghai (257 ml) and the highest beverages consumption in Shanghai (323 ml) and Beijing (264 ml). Consumption of tea in men (229 ml) was significantly higher than that in women (57 ml) (Z = 7.52, P = 0.000). Subjects in urban (693 ml) had lower consumption of plain water than those in rural (914 ml). The proportion was 32.4% (480/1483) for subjects with water drinking less than 1200 ml per day.

CONCLUSIONThe daily consumption of total drinking water, including plain water, tea and beverages is different in adults among different cities and is different in gender and regions. It is nearly 1/3 of subjects with daily total drinking water less than current Chinese recommended water intake (1200 ml).

Adolescent ; Adult ; Beverages ; China ; Drinking ; physiology ; Drinking Water ; Female ; Humans ; Male ; Middle Aged ; Seasons ; Urban Population ; Young Adult

ObjectiveTo analyze the high-frequency ultrasound image features of acute scrotum in children and explore the value of high-frequency ultrasonography in the diagnosis and differential diagnosis of the disease.

METHODSThis retrospective study included 256 children aged 2 days to 14 years undergoing color Doppler ultrasonography at 2 hours to 3 days after onset of acute scrotum. We analyzed the morphology, internal echo and blood supply of the testis in comparison with the clinical and pathological results.

RESULTSAmong the 256 cases, acute testicular torsion was found in 23, of which 16 were treated by complete resection the necrotic testis and the other 7 by surgical reduction of testicular torsion. Ultrasonographically, the involved testes presented different degrees of increase or decrease in volume, with uneven internal echoes, irregular hypoechoic flakes, and testicular hydrocele. Color Doppler flow imaging (CDFI) showed significant blood flow signals around the diseased testes but none within them. Acute testicular appendix torsion was found in 116 cases, in which ultrasonography manifested nodules with round or oval abnormal echoes between the upper pole of the testis and caput epididymidis, first hypoechoic and then gradually increased, heterogeneous internally. CDFI revealed enlarged epididymides and enriched testicular blood flow but no blood flow signals in the nodules. The 103 cases of acute epididymitis were ultrasonographically characterized by varied degrees of swelling of the involved epididymis with uneven internal echoes and rich blood flow signals on CDFI. Six of the cases were diagnosed as acute orchitis, with the ultrasonographic features of testicular swelling and low but uniform internal echoes, with rich blood flow signals on CDFI. Incarcerated inguinal hernia was confirmed in 15 cases, in which ultrasonography revealed intrusion of the hernia into the obviously enlarged scrotal sac with the mesentery and intestine in it, and blood flow visible on CDFI. Acute scrotal wall hematoma and edema was found in 8 cases, with the ultrasonographic characteristics of scrotal wall thickening, with visible blood flow signals on CDFI.

CONCLUSIONSHigh-frequency ultrasonography has a high sensitivity and specificity for acute scrotum in children, which can be applied as the first-choice clinical imaging modality and provide reliable evidence for the diagnosis and differential diagnosis of the disease.

OBJECTIVETo evaluate the efficacy and safety of the ICE regimen (iphosphamide + carboplatin + etoposide) used in treating children with hepatoblastoma in the Shanghai Children's Medical Center.

METHODSFrom June 2000 to June 2008, 14 children with newly diagnosed hepatoblastoma (7 males and 7 females) were enrolled. Their median age on diagnosis was 1.33 years (range: 0.25-8.25 years). Six patients had stage I disease, 1 had stage II, 5 had stage III, and 2 had stage IV diseases. Thirteen children had markedly increased serum AFP level, and 1 had normal serum AFP level. Multidisciplinary co-operation treatment was performed. Eight patients had primary surgery while 3 patients had pre-operation chemotherapy before surgery. ICE chemotherapy regimen was used. Totally, 73 courses of chemotherapy were administered for the 14 children and 25 out of the 73 courses were performed before operation.

RESULTSTwelve patients responded to the treatment (85.7%) and 2 failed. Ten patients (71.4%) achieved complete remission after treatment, and two had partial remission. By July 31st, 2008, 9 patients survived without any event, with a median follow-up duration of 35 months (range: 16-96 months). The 5-year overall survival rate was 70.71+/-12.37%, and the 5-year event-free survival rate was 64.29+/-12.81%. One patient had disease relapse and two patients were lost to follow-up after they achieved partial remission.

CONCLUSIONSThe ICE regimen combined with operation is effective and safe in treating children with hepatoblastoma.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carboplatin ; administration & dosage ; Child ; Child, Preschool ; Etoposide ; administration & dosage ; Female ; Hepatoblastoma ; drug therapy ; Humans ; Ifosfamide ; administration & dosage ; Infant ; Liver Neoplasms ; drug therapy ; Male

OBJECTIVETo examine the incidence and clinical significance of NPM1 mutations in childhood acute myeloid leukemia (AML) patients.

METHODSNPM1 mutations of 70 newly diagnosed childhood AML were detected by high resolution melting (HRM) analysis on the LightCycler 480. The incidence and clinical significance were analyzed.

RESULTSNPM1 mutations were identified in 32 (45.7%) of the 70 AML children. There was no significant difference in clinical characteristics between patients with or without NPM1 mutation, but patients with NPM1 mutation had a higher platelet count (P = 0.013). There was also no significant difference in NPM1 mutation between normal and abnormal karyotype groups. In AML-ETO or PML-RARα positive groups, the incidence of NPM1 mutations was significant lower (P = 0.048). There was no significant difference in response rates after induction therapy (P = 0.217), but the complete remission (CR) rate was higher in the NPM1-mutated group (81.3%). There was a trend toward higher event-free survival (EFS) and overall survival (OS) rates in the NPM1 mutated patients than that in wild NPM1 patients (EFS = 53.8% vs 41.4%, OS = 52.7% vs 39.2%), but the difference was not statistically significant (P = 0.374 and 0.380).

CONCLUSIONNPM1 mutations were relatively common in our cohort of AML patients. There was no significant difference in clinical characteristics between patients with and without NPM1 mutation. The NPM1 mutation patients group seemed to have better therapy response, but the difference was not statistically significant.

Child ; Disease-Free Survival ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Mutation ; Nuclear Proteins ; genetics ; Prognosis

OBJECTIVETo improve the long-term prognosis of childhood Hodgkin's lymphoma (HL) by standard treatment protocol HL-98.

METHODSPatients were divided into low (R(1)), middle (R(2)) and high-risk (R(3)) groups based on staging, tumor size and with or without B symptoms. Patients of R(1), R(2) and R(3) groups were given 4, 6, and 9 courses of chemotherapy, respectively. Low dose radiotherapy to involved area was given to patients with residual disease at the end of chemotherapy. All patients diagnosed between 1998 and Dec. 2008 were enrolled. The software of SPSS 11.0 was used and the event free survival (EFS) was generated by Kaplan-Meier.

RESULTSThere was a total of 26 patients with male 20 and female 6. The average age was 97 (30 to 179) months and median age 94.5 months. Three patients were in stage I, 4 in stage II, 9 in stage III and 10 in stage IV. Of 26 patients, 24 were found with neck tumor, 12 with mediastinum tumor, 11 with spleen infiltration and 5 with B symptom. Four patients were allocated into R(1) group, 12 R(2) group and 10 R(3) group. Eight of 26 with residual disease received radiotherapy, 7 received 20-26 Gy and 1 received 36 Gy. To Jun 2009, 21 (80.76%) of them kept in complete remission (CR) at 10 to 120 months follow-up (average 36 months, and median 31 months). Five cases relapsed (1 of stage III and 4 of stage IV) within 5 to 12 months. Three out of 4 in stage IV with B symptom relapsed. The estimated 5-year overall survival (OS) was 85.9% and EFS was 73.7%.

CONCLUSIONThe estimated 5-year EFS indicated that protocol HL-98 is reasonable good. Patients of stage I and II can obtain a good prognosis without radiotherapy.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Disease-Free Survival ; Follow-Up Studies ; Hodgkin Disease ; drug therapy ; Humans ; Neoplasm Staging ; Survival Rate