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

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

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.

Objective To study the effect of hesperetin(HSP)on renal ischemia-reperfusion injury(RIRI)in rats and its mechanism.Methods Fifty male SD rats were randomly divided into five groups:sham group,model group(renal ischemia-reperfusion injury I/R),HSP pretreatment(HSP+I/R)group,all-trans retinoic acid pretreatment(ATRA+I/R)group,HSP and ATRA pretreatment(HSP+ATRA+1/R)group.Each group had 10 rats.The sham group only exposed both kidneys by opening the abdominal cavity.In other groups,the RIRI model was established by occluding the bilateral renal pedicles for 45 min,followed by reperfusion for 24 h.Hematoxylin-eosin staining(HE)was used to ascertain the extent of kidney injury.The microplate method was used to measure serum creatinine(Scr)levels,while the urease method was used to measure blood urea nitrogen(BUN)levels.Enzyme-linked immunosorbent assay(ELISA)was used to detect the contents of interleukin-6(IL-6),interleukin-1β(IL-1 β)and tumor necrosis factor-α(TNF-α)in the serum.The kit method was used to examine the levels of superoxide dismutase(SOD),glutathione(GSH),malondialdehyde(MDA)and catalase(CAT)in kidney tissue.The Western blot analysis was conducted to detect the expression of nuclear factor erythroid 2 related factor 2(Nrf2)and its downstream antioxidant proteins heme oxygenase-1(HO-1)and quinone oxide reductase 1(NQO1)in renal tissues.Results The scores for renal tubule damage in sham,I/R and HSP+I/R groups were 0.20±0.45,4.20±0.84 and 2.40±0.55;Scr contents were(55.52±11.23),(207.10±19.22)and(105.60±18.11)μmol·L-1;IL-6 contents were(33.66±6.83),(172.50±8.09)and(105.40±10.03)pg·mL-1.The SOD levels in sham,I/R,HSP+I/R,ATRA+I/R and HSP+ATRA+I/R groups were(57.04±3.44),(37.29±3.60),(51.61±9.41),(32.55±5.58)and(37.40±3.66)U·mg·prot-1;the relative expression levels of Nrf2 were 0.37±0.24,0.57±0.28,1.31±0.34,0.44±0.17 and 0.77±0.25;the relative expression levels of HO-1 protein were 0.26±0.14,0.57±0.30,1.32±0.61,0.53±0.28 and 0.67±0.50.There was significant difference in renal tubule damage score,Scr,IL-6 and SOD indexes in I/R group compared to sham group(P＜0.05,P＜0.01).The above indexes were statistically significant between HSP+I/R group and I/R group(P＜0.05,P＜0.01).Conclusion The activation of HSP may stimulate the Nrf2-antioxidant response element(ARE)pathway,enhance the expression of downstream antioxidant proteins,mitigate the renal damage caused by oxidative stress,and ultimately improve RIRI.

Objective To explore the possibilities of human umbilical cord mesenchymal stem cells (HUC-MSCs) differentiate into vascular endothelial cells,thus to treat flap ischemic reperfusion injury and then promote the flap survival. Methods HUC-MSCs in vitro,and flow cytometric identification.Labeling stem cells with EdU,and then transplant stem cells into local of ischemiareperfusion injury flap of rats( treatment group),setting PBS control group.Observe the flap survival rate 7 days after,and execute rats.Take flap tissues for pathological sections,and detect vascular endothelial growth factor (VEGF) by immunohistochemical staining,detect the distribution or differentiation of the donor cells in cell receptor by EdU fluorescent staining.Results 7 days after,the flap survival rate of treatment group was ( 97.58 ± 3.41 ) ％,the flap survival rate of control group was (54.37 ± 8.78 ) ％,the rate of treatment group was higher than control group(P ＜0.05).The density of micrangium and VEGF of treatment group was higher than control group obviously ( P ＜ 0.05 ).The continuous distribution of positive cells that labeled with EdU were found in new vascular endothelial of treatment group' s survival flap tissues.Conclusions HUC-MSCs could differentiate into vascular endothelial cells,directly involved in generating new blood vesselsand,additionally could increase expression of VEGF to promote the formation of new blood vessels,then to set up the new micro circulation,accordingly to treat flap ischemic reperfusion injury and then promote the flap survival.

Objective To explore the possibilities of human umbilical cord mesenchymal stem cells (HUC-MSCs) differentiate into vascular endothelial cells,thus to treat flap ischemic reperfusion injury and then promote the flap survival. Methods HUC-MSCs in vitro,and flow cytometric identification.Labeling stem cells with EdU,and then transplant stem cells into local of ischemiareperfusion injury flap of rats( treatment group),setting PBS control group.Observe the flap survival rate 7 days after,and execute rats.Take flap tissues for pathological sections,and detect vascular endothelial growth factor (VEGF) by immunohistochemical staining,detect the distribution or differentiation of the donor cells in cell receptor by EdU fluorescent staining.Results 7 days after,the flap survival rate of treatment group was ( 97.58 ± 3.41 ) ％,the flap survival rate of control group was (54.37 ± 8.78 ) ％,the rate of treatment group was higher than control group(P ＜0.05).The density of micrangium and VEGF of treatment group was higher than control group obviously ( P ＜ 0.05 ).The continuous distribution of positive cells that labeled with EdU were found in new vascular endothelial of treatment group' s survival flap tissues.Conclusions HUC-MSCs could differentiate into vascular endothelial cells,directly involved in generating new blood vesselsand,additionally could increase expression of VEGF to promote the formation of new blood vessels,then to set up the new micro circulation,accordingly to treat flap ischemic reperfusion injury and then promote the flap survival.

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.