Objective To investigate the relationship between single nucleotide polymorphisms (SNPs) of PRKCG gene (rs2547362,rs3745406) and osteosarcoma susceptibility in the osteosarcoma patients and the normal population.Methods Sixtyone patients with osteosarcoma who had been admitted in our hospital from January 2011 to December 2012 and 63 healthy adults were enrolled in this study.A 2-ml peripheral blood sample was taken from each participant.The RT-qPCR method was used to detect the genotype and allele frequency distribution of PRKCG gene at rs2547362 and rs3745406 in osteosarcoma patients and normal population.Osteosarcoma patients were divided into several groups according to the clinical parameters such as age,gender,histology,tumor location,Enneking classification,tumor metastasis and therapy,and then we analyzed the relations between the genetic polymorphism and clinical parameters.Results 1) The genotype of PRKCG gene at rs3745406 included CC,CT and TT.The differences of genotypes (CC,CT,TF) and alleles (C,T) frequency distribution at rs3745406 were not statistically significant between osteosarcoma patients and the normal population (P=0.490,P=0.554).2) The genotype of PRKCG gene at rs2547362 included CC,CT and TT.The differences of genotypes (CC,CT,TT) and the alleles(C,T) frequency distribution at rs2547362 were statistically significant between the osteosarcoma patients and the normal population (P=0.006,P=0.007).3) The differences of genotypes (CC,CT,TT) and alleles (C,T) frequency distribution at rs3745406 were statistically significant between patients with metastasis and patients without metastasis (P=0.000,P=0.000).The CT and TT genotypes and the T allele carrier frequency at rs3745406 were higher in patients with metastasis than in patients without metastasis.SNPs at rs2547362 were not associated with clinical parameters.Conclusion The genetic polymorphism of PRKCG gene at rs2547362 is associated with osteosarcoma susceptibility.The TT genotype and T allele at rs3745406 are associated with metastasis of osteosarcoma,which may be a risk factor for metastasis in the osteosarcoma patients.

Objective To investigate the expression and clinical significance of P-selectin (CD62P) in bone marrow hematopoietic stem cells of patients with acute leukemia (AL). Methods The CD62P expression in bone marrow mononuclear cells of 15 healthy donors and 56 untreated patients with AL, were examined by flow cytometry. Results The average rate of CD62P expression was (6.72±7.64) % in hematopoietic stem cells (CD+45 CD+34 CD-38) of the 38 patients with acute myeloid leukemia (AML), was (3.46±2.51) % in hematopoietic stem cells (CD+45 CD+34 CD+19) of the 12 patients with B-acute lymphoblastic leukemia (B-ALL), and was (6.23±4.95) % in hematopoietic stem cells (CD+45 CD+34 CD+7) of 6 patients with T-acute lymphoblastic leukemia (T-ALL). The expression rates in those AL patients were higher than that in the healthy controls (1.04 ±1.23) % (t = 2.847, 3.284, 3.091, respectively, P ＜0.01), while there was no difference between the control group and the group who reach CR after routine treatment (t =1.932, P ＞0.05). Furthermore, the leukocyte,hemoglobin and platelet count in CD+62P patients with AML and T-ALL were significantly higher than CD-62P ones (t =4.153, 8.095, 8.289, 7.235, 8.692, 9.832, respectively, P ＜0.05), but there was no significant difference between CD+62P and CD-62P patients with B-ALL (t =0.340, 1.142, 0.019, respectively, P ＞0.05).Conclusion The CD62P is one of the markers of platelet activation, and its expression varies in different types of AL. The CD62P in hematopoietic stem cells of AL could be regarded as a new sign for the leukemic stem cells, as well as a helpful prognostic indicator in treatment response assessment.

According to ICH, Chinese Pharmacopoeia and supplementary requirements on the separation and purification of herbal extract with macroporous adsorption resin by SFDA, hexane, acetidine, ethanol, benzene, methyl-benzene, o-xylene, m-xylene, p-xylene, styrene, diethyl-benzene and divinyl-benzene of residual organic solvents and macroporous resin residues in Akebia saponin D were determined by headspace capillary GC. Eleven residues in Akebia saponin D were completely separated on DB-wax column, with FID detector, high purity nitrogen as the carry gases. The calibration curves were in good linearity (0.999 2-0.999 7). The reproducibility was good (RSD < 10%). The average recoveries were 80.0% -110%. The detection limit of each component was far lower than the limit concentration. The method is simple, reproducible, and can be used to determine the residual organic solvents and macroporous resin residues in Akebia saponin D.
Chromatography, Gas ; instrumentation ; methods ; Drug Contamination ; prevention & control ; Organic Chemicals ; analysis ; Reproducibility of Results ; Resins, Synthetic ; chemistry ; Saponins ; analysis ; isolation & purification

Transcranial magnetic stimulation (TMS) is a non-invasive diagnostic and therapeutic technigue. This paper expounds the design and manufacture of the TMS system, which meets all the requirements of the TMS study and clinical diagnosis and treatments.
Cerebral Cortex ; physiology ; Electric Stimulation ; instrumentation ; Electromagnetic Fields ; Equipment Design ; Humans ; Transcranial Magnetic Stimulation ; instrumentation

Objective To explore the constitute and antimicrobial susceptibility of pathogenic fungi causing candidemia in Nanchang City of Jiangxi Province.Methods Candida spp.isolated from blood specimens of patients at a hospital in Nanchang in March-October 2015 were collected, fungal strains were identified by amplifying the internal transcribed spacer (ITS) and large ribosomal subunit (D1/D2 region of 26rRNA), antifungal susceptibility of fungi was detected.Results A total of 1 332 positive blood culture specimens were collected, including 74 fungal positive specimens, accounting for 5.56％, 52 strains of Candida spp.were obtained, most were Candida tropicalis (n=17,32.69％),followed by Candida albicans(n=16, 30.77％) and Candida parapsilosis complex (n=16, 30.77％).Identification results of ITS and D1/D2 region were identical.52 strains of Candida spp.were sensitive to both micafungin and caspofungin, epidemiological cutoff value(ECV) of amphotericin B showed that 52 strains were all wild type.Resistance rates of Candida tropicalis to fluconazole and voriconazole were 29.41％ and 17.64％ respectively, ECV of itraconazole and posaconazole showed that wild type accounted for 82.35％ and 94.12％ respectively;resistance rates of Candida albicans to fluconazol and voriconazole were 93.75％ and 81.25％ respectively, ECV of itraconazole and posaconazole showed that wild type accounted for 75.00％ and 81.25％ respectively;Candida parapsilosis complex strains were sensitive to both fluconazole and voriconazole, ECV of itraconazole and posaconazole showed that all were wild type;all Candida glabrata strains had intermediate resistance rates to fluconazole, ECV of voriconazole, itraconazole, and posaconazole showed that wild type accounted for 66.67％, 100.00％, and 100.00％ respectively.Conclusion Candida tropicalis is the most common pathogenic fungus causing candidemia in Nanchang of Jiangxi, followed by Candida albicans and Candida parapsilosis complex.Azole, echinocandin, and amphotericin B are still first-line antifungal agents.

Objective To study the changes of PI3K/Akt/GSK3β signaling pathway during resuscitation with neck cooling in order to explore the relationship between the protective effect of neck cooling and the phosphorylation of PI3K/Akt and GSK3β.Methods Thirty rabbits were randomly(random number) divided into five groups, and models of cadiac arrest were induced by ventricular fibrillation(VF, the positive electrode in the right ventricle and negative pole on the apex of heart) for 4 min.In sham group,a electrode was placed into right ventricle without electric current conducted, and CA was not induced.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.In normothermia treat group(NT group),resuscitation was carried out to restoration of spontaneous circulation(ROSC),and the rabbits were sacrificed and specimens were taken at 24 hours after modeling.In intra-arrest therapeutic hypothermia group (IATH group), rapid neck cooling was initiated at the same time with CPR,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.Rabbits were sacrificed and specimens were taken at 24 hours after modeling.In recovery period cooling + LY294002 group(PATH+LY294002 group), LY294002 was injected intra-ventricularly at 20 minutes before resuscitation.Rapid neck cooling was started at the same time with CPR,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.In post-arrest therapeutic hypothermia group (PATH group), rapid neck cooling was begun after CPR for 1 hour,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.Animals were sacrificed by using overdose anesthetic drug.Western blot was used to detect the level of Akt p-Akt GSK-3β p-GSK-3β (ser9) protein, and TUNEL was used to observe apoptosis of tissues in each group.Multiple comparisons were performed with one-way analysis of variance (ANOVA).Results Compared with Sham group, Akt (Thr-308) phosphorylation (P-AKT) and P-GSK-3β levels in the brain neuron cytoplasm in 24 hours after CPR resuscitation in NT group was significantly reduced, and showed a gradual reduction trend (P<0.05);the P-AKT and P-GSK-3β levels in the brain neuron cytoplasm in 24 hours after CPR resuscitation in IATH group were significantly enhanced compared with NT group (P<0.05);the levels of these two kinds of protein at one hour after resuscitation in PATH group were significantly enhanced compared with NT group (P<0.05), but lower in IATH group.Intra-ventricularly injection of LY294002 made the effect of hypothermia lost, indicating that LY294002 inhibited the phosphorylation of Akt.Apoptosis cells were significantly reduced in IATH group and normothermia theatment group compared with PATH group and LY294002 group(P<0.05).Conclusions Neck cooling can reduce apoptosis in rabbit brain cells after recovery, and the protective effect on brain is best in intra-arrest therapeutic hypothermia group.LY294002 specifically block the PI3K/Akt pathway, and the protective effect of cooling on the brain can be abolished,indicating hypothermia protects the neurological function via activation of PI3K/Akt pathway.Neck cooling protects the neurological function by activating PI3K/Akt/GSK-3β, promoting the Akt activation, and increasing the expression of P-GSK3β.Specific Akt inhibitor LY294002 inhibits Akt phosphorylation of brain tissue recovery and further inhibit the phosphorylation of GSK-3β, thus abolishing protective effect of cooling on neurological function.

Objective To observe the expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblast,and to study the role of fibronectin in pathogenesis of chronic fluorosis.Methods Male and female Wistar rats 144 were randomly divided into four groups,which were designated as the control group(normal diets,n =36),fluoride group(normal diets + 100 mg/L fluoride,n =36),lower calcium monophagia group (synthetic diets,n =36) and lower calcium monOphagia with fluoride group(synthetic diets + 100 mg/L fluoride,n =36).Rats were sacrificed 4 and 8 months after beginning of the experiment,respectively,and femur tissue was fixated and paraffin-embedded.The osteoblast isolated from calvaria of neonatal rats was treated with different dose of fluoride(0,1,2,4 mg/L fluoride,respectively) for 48 and 72 h,cell culture supernatant and cells were collected,respectively.The cranial osteoblasts were cultured in vitro and divided into four groups according to different concentration of fluoride added,which were 0(control group),0.01,1.00,and 10.00 mg/L groups.These cells were treated with mineralized induced medium at day 2 and cultured for 3 more weeks whereafter,and then the slides were fixed in alcohol.The expression of fibronectin in rat femur tissue was detected by immunohistochemistry (IHC),and fibronectin mRNA expression was determined by in situ hybridization; the fibronectin levels in supernatant of cultured osteoblast was examined by enzyme-linked immunosorbent assay(ELISA),and the expression of fibronectin mRNA in osteoblasts was detected with RT-PCR; skull mineralized nodule formation of osteoblasts was observed under a light microscopy after stained with 0.1％ red alizarin liquid.Results Little expression of fibronectin (brown granules under light microscope) could be seen in femur tissue of fluorosis rats of control group and lower calcium monophagia group; but abundantly expressed in fluoride group and lower calcium monophagia with fluoride group; the fibronectin was also expressed in osteoblasts,bone cells and bone marrow cells with less red particles in the control group and lower calcium monophagia group,but more in the fluoride group and lower calcium monophagia with fluoride group.The expression of fibronectin protein in supernatant of cultured osteoblasts was significantly increased in the group of 4 mg/L fluoride at 48 h(0.108 ± 0.042,t =0.764,P＜ 0.05) compared with control group(0.081 ± 0.010); the value was also significantly increased in 1,2,4 mg/L groups at 72 h(0.089 ± 0.010,0.087 ± 0.012,0.098 ± 0.023; t =0.765,0.704,0.996; all P ＜ 0.05) compared with control group (0.070 ± 0.014) ; the expression of fibronectin mRNA was much higher in 1,2,4 mg/L groups at 48 h (0.61 ±0.06,0.77 ± 0.07,0.77 ± 0.07) and 72 h(1.61 ± 0.14,2.54 ± 0.20,2.75 ± 0.22) compared with control group [0.48 ± 0.04(t =0.111,0.182,0.182,all P ＜ 0.05),0.97 ± 0.08(t =0.093,0.109,0.108,all P＜ 0.05) ].A lot of mineralized nodules could be seen under light microscope in 1.00 and 10.00 mg/L groups.Conclusions The expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblasts are increased,and fluoride also promotes the mineralization nodules formation of osteoblasts.These results suggest that fibronectin may regulate the process of bone mineralization,and possibly play a role in the development of skeletal fluorosis.

·AIM: To detect the effect of CTGF on the apoptosis in the diabetic retina with small interfering RNAs (siRNA) targeting with CTGF. ·METHODS: A total of 60 rats were divided into six groups including control group, diabetic 4,8,12,16 weeks group, and interference group. Diabetic rats were induced by STZ intra-peritoneal. At 4, 8, 12, 16 weeks after diabetic setting up, retinas were obtained from control, diabetic rats and diabetic animals treated by intravitreal injection of CTGFsiRNA to suppress the expression of CTGF mRNA. Retinal cells apoptosis was detected by Tunnel staining and mRNA expression of CTGF was analyzed by RT-PCR.·RESULTS: The levels of CTGF and the apoptosis in the retinas of diabetic rats were significantly higher than those in the controls. Apoptosis occurred at 4 weeks after a diabetic model setting up, became serious with the diabetes developing, while CTGF elevated at 8 weeks. The cell apoptosis counts increased to 25.8cells/mm2 at 24 weeks of diabetes. SiRNA-mediated inhibition of CTGF mRNA resulted in a significant decrease in apoptosis. Significant correlations were found between CTGF and apoptosis in the retina.·CONCLUSION: These results suggest that CTGF might be involved in retinal cells apoptosis which is a characteristic of early diabetic retina. siRNA targeting CTGF seems to have the advantage of ameliorating retinal cells lost.

OBJECTIVETo explore the feasibility of using transpedicular balloon kyphoplasty for aged osteoporotic thoracolumbar burst fractures with an in vitro model.

METHODSSimulated osteoporotic thoracolumbar burst fractures were created in 11 vertebral bodies. The burst fractures without obvious canal occupation were confirmed by spiral CT before the procedure. This operation involved the percutaneous insertion of two inflatable bone tamps into a fractured vertebral body transpedicularly under fluoroscopic guidance. Inflation of the bone tamp elevated the endplates, restored the vertebral body height, while created a cavity to be filled with bone cement. Preoperative and postoperative heights were measured, preoperative and postoperative sagittal diameter of vertebral canals were measured, and the repaired vertebral bodies were compressed to determine strength and stiffness values. The cement distribution and extravasation was evaluated by spiral CT after the procedure.

RESULTSThe transpedicular balloon kyphoplasty resulted in the significant restoration of the vertebral body height. The mean vertebral body heights was (20.73 +/- 1.37) mm before operation and (24.82 +/- 1.61) mm after operation (P < 0.01). Preoperative and postoperative sagittal diameter of vertebral canals were similar (P > 0.05). The cement distribution was regular, and the extravasation into spinal cannula was found in one case. The preoperative strength was significantly larger than the postoperative strength, whereas the postoperative stiffness was significantly less than the preoperative stiffness (P < 0.05).

CONCLUSIONSTranspedicular balloon kyphoplasty is an effective way to treat aged osteoporotic thoracolumbar burst fractures without obvious canal occupations and neurologic deficit in vitro. The procedure can increase strength and restore height of aged osteoporotic thoracolumbar burst fractures. Cement distribution and vertebral canal stenosis can be avoided.

Cadaver ; Humans ; Kyphoplasty ; instrumentation ; methods ; Kyphosis ; diagnostic imaging ; etiology ; surgery ; Osteoporosis ; complications ; Spinal Fractures ; complications ; diagnostic imaging ; surgery ; Spinal Fusion ; methods ; Thoracic Vertebrae ; pathology ; surgery ; Tomography, X-Ray Computed

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