Objective T o detect the genotype of A B O blood group by SN aPshot technology. Methods D N A w ere extracted from the peripheral blood sam ples w ith know n blood groups (obtained by serology) of 107 unrelated individuals in Y unnan. Six SN P loci of the 261th, 297th, 681th, 703th, 802th, and 803th nucleotide positions w ere detected by SN aPshot M ultiplex kit, and relevant genetics param eters w ere cal-culated. Results In 107 blood sam ples, the allele frequencies of types A , B , O A, and O G w ere 0.3551, 0.1682, 0.2300 and 0.2476, respectively, w hile that of types A G and cis A B w ere not detected. T he geno-typing results of A B O blood group w ere consistent w ith that of serologic testing. Conclusion SN aPshot technology can be adapted for genotyping of A B O blood group.

Oroxylum indicum was a traditional Chinese medicine. In order to study the chemical constituents from the seed of O. indicum, the chemical constituents of 80% methanol extract of seeds of O. indicum were subjected to chromatography on silica gel, Sephadex LH-20, and preparative HPLC, leading to the isolation of eleven compounds. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR and 13C-NMR data as oroxin B (1), chrysin (2), baicalein (3), neglectein (4), quercetin-3-O-β-D-galactopy ranoside (5), quercetin-7-O-β-D-glucopyranoside (6), 2α,3β-dihydroxylluPeol (7), lupeol (8), rengyol (9), β-sitostero (10), and stigmasterol (11). Among them, compound 5 were firstly obtained from O. indicum.
Bignoniaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Seeds ; chemistry

BACKGROUNDNowadays it is now a focus topic in schistosomiasis research to find ideal vaccine candidates and new drug targets for developing anti-schistosomiasis vaccine. We cloned a new gene, casein kinase II beta subunit, of Schistosoma japonicum (S. japonicum) and express it in Escherichia coli (E. coli).

METHODSThe ESTs obtained in our laboratory were analyzed by homologous searching, and a new gene was recognized. The full-length cDNA of the new gene was obtained by joining the 3'RACE PCR fragment and the EST clone. To express the new gene, the cDNA was cloned into pGEX-4T-1 vector and then transformed into E. coli JM109. The recombinant protein was analyzed by SDS-PAGE and Western-blot.

RESULTSA 908 bp cDNA was isolated from S. japonicum and identified to be casein kinase II beta subunit gene by sequence analysis. The open reading frame of the gene encodes a protein of 217 amino acids exhibiting 75.8%, 75.8%, 73.9%, 68.2%, 51.6% identity to the amino acids sequence of the corresponding genes of Homo sapiens (H. sapiens), Xenopus laevi (X. laevi), Drosophila melanogaster (D. melanogaster), Caenorhabditis elegan (C. elegan), and Schizosaccharomyces pombe (S. promber) respectively. The predicted molecular weight of the protein was 24.921 kDa. The new cDNA sequence had been submitted to GenBank, and its accession number is AY241391. This cDNA was subcloned into the pGEX-4T-1 vector and expressed in E. coli JM109. The recombinant protein could be recognized by the S. japonicum infected rabbit serum.

CONCLUSIONThe full-length cDNA sequences encoding S. japonicum casein kinase II beta subunit were firstly sequenced, cloned, and expressed in E. coli.

Amino Acid Sequence ; Animals ; Base Sequence ; Blotting, Western ; Casein Kinase II ; chemistry ; genetics ; Cloning, Molecular ; DNA, Complementary ; chemistry ; isolation & purification ; Escherichia coli ; genetics ; Molecular Sequence Data ; Rabbits ; Schistosoma japonicum ; enzymology ; genetics

Objective To investigate selcnium(Se) levels of environment and human body in Gaomi City and Zichuan District of Shandong. Methods Lijiaying Township in Gaomi City of Weifang City, Zhaili Township and Longquan Township in Zichuan District of Zibo City were selected. Two farming soil samples at different spot, local wheat and corn, residents nail samples from 3 to 4 families were collected in each natural village in the investigated towns. The contents of Se were detected by 2,3-diamino naphthalene fluorescence method. Results Se level of the soil, wheat, corn, and nails in Lijiaying [(0.054 ± 0.019), (0.022 ± 0.009), (0.018 ± 0.007), (0.365 ± 0.108)mg/kg] was significantly lower than that in Zhaili [(0.425 ± 0.080), (0.130 ± 0.043), (0.098 ± 0.026), (0.751 ± 0.134)mg/kg] and Longquan[(0.487 ± 0.153), (0.112 ± 0.030), (0.097 ± 0.029), (0.735 ± 0.145)mg/kg;P < 0.01]. In Lijiaying, Se was deficient in soil, wheat, corn(< 0.200, < 0.025 mg/kg), above Se deficiency diagnosis and below Se-adequate level in the nail, while in Zhaili and Longquan, the Se level in the soil (0.425, 0.487 mg/kg), wheat(0.130, 0.112 mg/kg), corn (0.098, 0.097 mg/kg), nails (0.751, 0.735 mg/kg) was adequate (≥0.400 mg/kg). Conclusions The external environment is Se-deficient in Lijiaying, Se-adequate in Longquan and Zhaili. The selenium level in human body is consistent with the external environment.

Animals ; Immunohistochemistry ; In Situ Hybridization ; Liver ; enzymology ; pathology ; Liver Cirrhosis ; enzymology ; etiology ; pathology ; Male ; Matrix Metalloproteinase 2 ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar

OBJECTIVETo explore the effects of Linggui Zhugan Decoction (LZD) combined calorie restriction on fasting plasma glucose (FPG), the insulin resistance (IR), and the peroxisome proliferator-activated receptor gamma (PPAR-gamma) of IR model rats.

METHODSTotally 48 male Wistar rats were randomly divided into the control group, the model group, the calorie restriction group, and the TCM + calorie restriction group, 12 in each group. Ordinary forage was given to those in the control group, and high fat diet was fed to those in the rest 3 groups for 12 weeks to establish the IR model. After successful modeling, rats in the control group and the model group were continually fed with the original farage for 4 days. The normal saline at the daily dose of 20 mL/kg was given to them by gastrogavage. The normal saline at the daily dose of 20 mL/kg was given to rats in the calorie restriction group by gastrogavage after 4-day calorie restriction. LZD at the daily dose of 20 mL/kg was given to rats in the TCM +calorie restriction group by gastrogavage after 4-day calorie restriction. The body weight, FPG, serum fasting insulin (FINS), insulin resistance index (IRI), and the protein expression of PPAR-y in the omental adipose tissue were compared.

RESULTSAfter 4-day calorie restriction, the body weight obviously decreased in the calorie restriction group and the TCM +calorie restriction group, when compared with the model group (P <0.01). There was no statistical difference between the former two groups (P >0.05). The FINS and IRI obviously decreased in the calorie restriction group (P <0.01, P <0.05). The FPG, FINS, and IRI significantly decreased in the TCM + calorie restriction group (P <0. 05, P <0.01). The protein expression of PPAR-gamma obviously decreased in the calorie restriction group and the TCM + calorie restriction group (P <0.01).The phlegm dampness state was alleviated, with more significant effects shown in the TCM + calorie restriction group.

CONCLUSIONSLZD combined calorie restriction could reduce the body weight, FPG, and IRI of IR rats. Besides, it showed better effects than calorie restriction alone. Its effects in improving IR might be correlated with inhibiting the activities of PPAR-gamma. Meanwhile, it might play a role in inhibiting the differentiation of fat cells.

Animals ; Blood Glucose ; analysis ; Caloric Restriction ; Drugs, Chinese Herbal ; pharmacology ; Insulin ; metabolism ; Insulin Resistance ; Male ; PPAR gamma ; metabolism ; Rats ; Rats, Wistar

Objective: To explore the application of the 3D precise digital technology in restoring structural defects of the orbital region. Methods: Structural defects of the orbital region concerning on osseous structure and soft tissue were restored in one stage or stages using EH composite artificial bone prosthesis with complex three-dimensional structure. The fabrication of EH composite artificial bone prosthesis was based on CT scanning and 3D reconstruction of the skull with computer aided data analysis and design. The appearance, degree of satisfaction and the complications were evaluated in postoperative regular follow-up. Results: Five cases of structural defects in the orbital regions presenting bone defect and soft tissue abnormality, received treatments in the department from January 2016 to September 2017. The cases consist of one patient with dysplasia following surgical treatment, three with post-traumatic and one with Treacher-Collins syndrome. With the application of individualized EH composite artificial bone fabricated by aforementioned method, all the repair materials presented the ideal three-dimensional structure and coincided well with the defects, and soft tissue restoration of 2 cases was performed in one stage or by stages. Appearance and symmetry of the 5 cases was significantly improved, without complications of infection, rejection, exposure or graft tissue necrosis. All the patients were satisfied with the results. Conclusions In consideration of the capacity to fabricate accurate individualized repair materials, the three-dimensional digital technology plays an important role in the treatment of structural defects in the orbital region, especially the reconstruction of the bony contour. The simple orbital deformities can be treated with the repair materials and correction of soft tissue. For special orbital deformities, attention should be paid to bone structure repair, eye socket reconstruction and filling of orbital contents sequentially.

OBJECTIVETo study the changes of serum total calcium (Ca) and inorganic phosphate (P) levels during children growth and related factors.

METHODSUsing a stratified-multi-steps-cluster sampling method, we selected 2,342 healthy children aged 10-18 years from urban and suburban areas of Beijing from September 2001 to December 2001 to test the levels of serum Ca and P. Their meal status was also surveyed to analyse the correlation of the leves of serum Ca and P with sex, age, height, weight, and diet on statistic basis.

RESULTSThe Ca levels of Beijing children were (2.39 +/- 0.12) mmol/L, which had a positive correlation with age, height, and weight (P < 0.01). The Ca levels of female children were somewhat higher than those of male [male, (2.38 +/- 0.12) mmol/L; female, (2.39 +/- 0.12) mmol/L; P < 0.05]. The Ca levels of urban children were significantly higher than those of suburban children [urban, (2.40 +/- 0.13) mmol/L; suburban, (2.37 +/- 0.10) mmol/L; P < 0.001]. The P levels of Beijing children were (1.39 +/- 0.18) mmol/L, which had a negative correlation with age, height, and weight (P < 0.01). The P levels of male children were significantly higher than those of female [male, (1.43 +/- 0.18) mmol/L; female, (1.36 +/- 0.17) mmol/L; P < 0.001]. The P levels of urban children were significant higher than those of suburban children [urban, (1.41 +/- 0.19) mmol/L; suburban, (1.38 +/- 0.16) mmol/L; P < 0.001]. The Ca levels of Beijing children had a negative correlation with P levels (r=-0.141, P < 0.01). [Ca] x [P] (mmol/L) of Beijing children were 3.32 +/- 0.44. The value of [Ca] x [P] reached peak by 3.45 +/- 0.46 when Beijing children were of 13-14 years old, and then the value declined with increasing age.

CONCLUSIONThe levels of serum Ca and P correlates with sex, age, growth, and diet. The level of serum Ca goes up while P goes down during the children growth.

Adolescent ; Age Factors ; Body Height ; Body Mass Index ; Body Weight ; Calcium ; blood ; Child ; Child Development ; Female ; Humans ; Male ; Phosphorus ; blood ; Reference Values ; Sampling Studies ; Sex Factors ; Urban Health

Objective:To prepare the synaptophysin and chromogranin A monoclonal antibodies with clinical evalua -tion.Methods:The fuse gene (Syn and CgA) was designed and it was constructed on the expression vector pET-28a.Then ,the fusion protein was purified.After protein immunization , cell fusion and screening , the target antibodies were selected .Specificity study and correlation coefficient of Syn and CgA was evaluated by clinical sample comparison validation .Results:By screening,two antibodies 3D9 and 4A12,respectively,for Syn and CgA,were obtained.19 kinds of wax block organization were detected by 3D9,4A12 and control antibody(Leica).The statistical results were analyzed ,the results were in good agreement ,and the correlation coefficients were r=0.9892 and r =0.9939, respectively.Conclusion: This method is prepared to obtain the synaptophysin and chromogranin A antibodies successfully and both can be used for immunohistochemistry .This method can also provide some reference for the study of antibody .

Objective To investigate the safety and acute toxicities of intraoperative electron radiotherapy for patients with abdominal tumors.Methods From May 2008 to August 2009, 52 patients with abdominal tumors were treated with intraoperative electron radiotherapy,including 14 patients with breast cancer,19 with pancreatic cancer,3 with cervical cancer, 4 with ovarian cancer, 6 with sarcoma, and 6 with other tumors.Fifteen patients were with recurrent tumors.The intraoperative radiotherapy was performed using Mobetron mobile electron accelerator, with total dose of 9 - 18 Gy.In all, 29, 4 and 19 patients received complete resection, palliative resection and surgical exploration, respectively.The complications during the operations and within 6 months after operations were graded according to Common Terminology Criteria for Adverse Events v3.0 (CTC 3.0).Results The median duration of surgery was 190 minutes.Intraoperative complications were observed in 5 patients, including 3 with hemorrhage, 1 with hypotension,and 1 with hypoxemia, all of which were treated conservatively.The median hospitalization time and time to take out stitches was 12 and 13 days, respectively.And the in-hospital mortality was 4% (2/52).Twentyfour patients suffered post-operative adverse events, including 3 postoperative infections.With a median follow-up time of 183 days, 20% of patients sufferred from grade 3 to 5 adverse events, with hematological toxicities being the most common complication, followed by bellyache.Grade 1 and 2 toxicities which were definitely associated with intraoperative radiotherapy was 28% and 4%, respectively.None of grade 3 to 5 complications were proved to be caused by intraoperative radiotherapy.Conclusions Intraoperative electron radiotherapy is well tolerable and could be widely used for patients with abdominal tumors, with a little longer time to take out stitches but without more morbidities and toxicities compared surgery alone.