Objective To discuss the clinical characteristics and treatment methods of lumbar disc herniation in adolescent and to in-crease the awareness of lumbar disc herniation among orthopedists.Methods Retrospectively reviewed the clinical data of 201 adolescent pa-tients(aged 20 years or less)with lumbar disc herniation who were admitted in General Hospital of Shenyang Military Area Command from January 2001 to December 2010.Analyzed the clinical characteristics and treatment methods,and compared the VAS score and ODI score be-fore operation,one week after operation and at the last follow up.Meanwhile,the complications were observed and compared.Results Among the 201 patients,187 patients(93.0%)presented with low back pain with or without radiating pain,20 patients(10.0%)presented with leg pain for the first symptom.There were only 97 patients(48.3%)were diagnosed with lumbar disc herniation for their first diagnosis in other hospital.Totally 70 patients(34.8%)had a history of trauma before the onset.The most common segments were L4 /L5 (101 cases,50.2%) and L5 /S1 (70 cases,34.8%).There were 140 patients treated by microendoscopy discectomy(MED),25 patients treated by percutaneous endoscopic lumbar discectomy(PELD),and 36 patients treated by open lumbar discectomy(OLD).All the patients were followed up for more than 12 months with averagely followed up for(22.9 ±11.7)months.There was significant improvement one week after operation and at the last follow up compared with the preoperative VAS score and ODI score,and the differences were statistically significant(P <0.05).The rate of postoperative complications was 3.6%,4.0%,8.3% in patients with MED,PELD and OLD respectively.Conclusion Diagnosis of LDH in adolescent is usually delayed.Increase the awareness of lumbar disc herniation could help orthopedists to get a exact medical history,per-form a directed physical examination,and order appropriate imaging studies.Minimally invasive surgery is an effective method to treat lumbar disc herniation in adolescent.

Developmental changes in children can affect drug disposition and clinical effects. A physiologically-based pharmacokinetic (PBPK) model is a mathematical model that can be used to predict blood drug concentrations in children and gain insight into age-dependent physiological differences in drug disposition impact. Pediatric PBPK (P-PBPK) models have attracted attention over the past decade. With the concerted efforts of academia, pharmaceutical companies, and regulatory agencies, there are more and more examples of pediatric clinical studies using PBPK models. Nevertheless, the number of P-PBPK models and their predictive performance still lag behind adult models. By referring to the literature, we study the process of children adapting to adult absorption, distribution, metabolism, and excretion (ADME) parameters and analyze the general principles of P-PBPK model establishment. In addition, we summarize the functions and application examples of commonly used P-PBPK modeling software to provide a basis for the rational application of modeling software.

Objective To confirm the death of a child injured by a dog was due to rabies and to understand the molecular biologic features of rabies virus in Kaili,Guizhou province.Methods Brain tissue samples of patient and dog were collected to detect the rabies virus by direct immunofluorescence assay (DFA) and RT-nested PCR assay.Homology and phylogenetic tree were analyzed based on the whole nucleotide and deduced amino acid sequence of N gene of rabies virus followed by molecular epidemiological analysis.Results Both the human and dog brain tissue samples were confirmed positive by DFA and RT-nested PCR assay.The homology analysis of N gene sequences among GZH,GZD and other epidemic and vaccine rabies strains isolated from other provinces and other countries indicated that the detected samples shared the highest homology with the strain detected in Anlong prefecture in Guizhou in the year of 2006,and the homology between GZH and GZD was as high as 100％.Besides,among the vaccine strains,GZH and GZD showed the highest homology with strain CNT.Phylogenetic analysis indicated that the two samples were very close and belonged to genetype 1 lyssavirus,with the closest relationship between samples reported in Guizhou and Beijing.Conclusion It was confirmed on the viral molecular level that both the human and dog in Kaili were suffered from rabies,and the pathogens were genetype 1 lyssavirus.The prevalent strains in Kaili city was probably imported from other prefectures of Guizhou province,suggesting that prevention and control measures on rabies in Guizhou province should be strengthened.

To study the characteristic of glycoprotein gene sequence of rabies virus in Guizhou Province in recent years and provide scientific basis for effective control and prevention of rabies,RT-PCR assay were used to detect human and dog brain tissues derived from different prefectures of Guizhou.The amplification products were sequenced and analyzed with bioinformatics software.The results showed that the full-length G gene sequences of 8 positive samples were obtained by RT-PCR amplification,sequencing and splicing.The homology of eight G gene sequences from Guizhou Province were between 87.4％ -99.9％ and 83.3.％- 100％ on nucleotide and deduced amino acid level,respectively,and the highest homology were found with the genotype 1 strains ( 86.5 ％- 87.0％ for nucleotide and 83.3 ％- 100 ％ for amino acid) among genotype 1- 7 representative strains.Besides,phylogenetic analysis based on the G gene indicated that the relationship of 8 strains derived from Guizhou were closest to genotype 1 Lyssavirus,and the strains of Guizhou were very close to the strains come from Hubei,Hunan,Anhui,Guangxi,Jiangsu provinces and Shanghai,except for GZ01 and GZ09.Moreover,GZ09 were evolutionarily closed to the strains come from Malaysia and Thailand,while the remaining sequences were closed to the strain of Indonesia.This study confirmed on the G gene level that rabies virus epidemic strains circulated in Guizhou Province in recent years belonged to rabies virus genotype 1 and the evolutionary relationship with reported strains come from different provinces of China and different countries were revealed in this study.It will provide scientific basis for effective control and prevention of human and animal rabies in Guizhou Province.

ObjectiveTo evaluate in vivo antithrombosis property of optimized FW-Ⅱ axial blood pump and provides evidence for future clinical use.MethodsA left ventricle-pump-descending aorta bypass model was established in five healthy sheep (60-70 kg) and the circulation of these sheep was assisted by FW-Ⅱ axial blood pump for 2 weeks.In preoperative and postoperative day 1,2,3,7,10 and 14,blood was drawn from the jugular vein to examine platelet activation and leukocyte-platelet aggregation respectively quantified with Annexin V,CD41/61 and CD14-PE by flow cytometry assays.Immediately after termination of the experiment,FW-Ⅱ axial blood pumps were explanted and each part was inspected for thrombus formation.Macroscopic and histological examinations were checked on heart,brain,kidney and spleen,respectively for thrombosis.ResultsCompared with preoperative baseline,the number of platelet activation and leukocyte-platelet aggregation reached a peak at postoperative day 2,it retained a high level within 7 days,then gradually decreased,but was still higher than preoperative level at dayl4.According to rotating speed,the number of platelet activation and platelet-leukocyte aggregation were lowest at the speed of 8000 r/min Minus thrombus were found in the front and rear hub of the pump rotor,and there was no thrombus at other components (flow straighter,impeller and pump housing).There were no ischemia and infarction evidences in macroscopic and histological examination of the heart,brain,kidney and spleen.ConclusionFW-II axial blood pump can be used to assist left ventricular circulation for 2 weeks with a satisfactory antithrombosis property.The level of platelet activation and leukocyte-platelet aggregation can be reduced to a lowest level at an optimized pump rotating speed.

Objective To investigate the combined effect and mechanism of metformin (Met) and 2-deoxy-D-glucose (2DG) on cell proliferation and apoptosis in liver cancer cells HepG2 and Hep3B.Methods Wst-1 reagent was used to determine the anti-proliferation effects after treatments with Met and 2DG alone or combined in HepG2 and Hep3B cells.Microscopy was used to observe cell morphological changes after treatments with Met and 2DG alone or combined in HepG2 and Hep3B cells.Cell apoptosis was observed by flow cytometry after treatment of different kinds of drugs.Western blotting was used to analyze the protein expressions of Caspase-3,PARP,Mcl-1 of HepG2.Results The survival rate of HepG2 cells in the combination group was (22.48 ± 0.51)％,and compared with the control group (100.00 ± 5.05)％,Met group (80.68 ±5.10)％ and 2DG group (72.56 ±4.34)％,the differences were statistically significant (P ＜ 0.001;P ＜ 0.001;P =0.001).The survival rate of Hep3B cells in the combination group was (29.16 ± 1.34) ％,and compared with the control group (100.00 ± 1.23) ％,Met group (59.58 ± 1.92) ％ and 2DG group (33.87 ± 1.95) ％,the differences were statistically significant (P ＜ 0.001;P ＜ 0.001;P =0.001).Microscopy observation showed that combined treatment of Met and 2DG caused less viable adherent cells of HepG2,but more floating dead cells.While the combination group also caused a decrease in the density of Hep3B cells,but did not significantly increase the shedding of cells.The apoptosis of HepG2 cells in the combination group was (39.63 ± 0.21) ％,and compared with the control group (7.12 ± 0.14) ％,Met group (12.56 ± 0.35) ％ and 2DG group (15.16 ± 1.93) ％,the differences were statistically significant (P ＜0.001;P ＜ 0.001;P =0.001).The apoptosis of Hep3B cells in the combination group was (12.58 ± 1.03) ％,and compared with the control group (2.82 ± 0.51) ％ and Met group (8.98 ± 0.86) ％,the differences were statistically significant (P ＜ 0.001;P =0.007),but compared with the 2DG group (12.40 ± 1.78) ％,the difference was not statistically significant (P =1.000).Furthermore,Western blotting demonstrated that the combined treatment induced evident Caspase-3 activation and poly ADP-ribose polymerase (PARP) cleavages,and decreased expression of Mcl-1.Conclusion The combination of Met and 2DG can effectively inhibit cell proliferation of HepG2 and Hep3B,and induce apoptosis of HepG2 cells.The mechanism may be involved with Caspase-3 activation,cutting PARP substrate and decreasing Mcl-1 protein.

Objective To investigate the role of CD4 ~+ CD25~+ regulatory T lymphocytes (Treg)in modulating the cellular immune response and pathogenesis of murine pulmonary tuberculosis.Methods Inactivation of Treg was achieved by intraperitoneal injection anti-CD25 (clone PC61,50 μ/mouse) in PC61 group, and rat-IgG (50 μ/mouse) was injected intraperitoneally in control group. All the mice were inoculated intravenously with H37Rv 0. 1 mL (1 × 10~6 CFU) 3 days after Treg inactivation. The effects of Treg inactivation in different tissues were analyzed by flow cytometry. The cellular immune response, pulmonary histopathology and bacterial load were determined in vitro at different time points. The data were compared using homogeneity of variance F test and non-paired t test. Results In spleen, the percentages of Treg/CD4 T lymphocytes in PC61 group and control group were (21. 13± 3. 58)% and (30. 42± 4. 20)%, respectively at day 10 of inoculation (t = 2. 38, P < 0. 05), and those were (16. 12 ± 1. 26)% and ( 17. 34± 1. 62)%,respectively at day 30 of inoculation (t = 0. 84,P>0. 05). The percentages of Foxp3~+/CD4~+ T lymphocytes in PC61 group and control group were (32. 07 ± 3. 95)% and (60. 55 ± 5. 48)%,respectively at day 10 of inoculation (t = 5. 96, P<0. 05). Similar results were achieved in the peripheral blood. Bacillus calmette-guerin (BCG)-specific 1L-17 (ng/L) secreted by murine spleen cells in PC61 group and control group at day 10, 30 and 60 of inoculation were 5. 1± 0.9 vs 0, 43. 1± 10.0 vs5. 9± 2. 8 and 124.8 ± 5.8 vs 102. 5±8. 1, respectively (t = 7. 90, t=5. 10,t = 3. 19; all P<0.05); those of BCG-specific IFN-γ (ng/L) were 28. 4 ± 8. 2 vs 4. 0±1. 3, 685. 9± 128. 6 vs418. 7±20.4 and 310.9±119. 7 vs 32. 8±7. 5, respectively(tO = 4. 21,t = 8. 43, t = 3. 27; all P<0.05);those of TNF-a (ng/L) were 38. 6±5.0 vs 16. 3±4. 0, 112. 9 ±12. 3 vs 71. 5±12. 6 and 86. 2±8. 2vs0, respectively(t = 4. 95, t=3. 33,t/=14.8; all P<0. 05). The lung bacterial load at day 10 of inoculation in PC61 group was lower than that in control group (t = 4. 63, P < 0. 01), but the differences were not significant thereafter. The changes of lung histopathology at late stage of infection (day 120) in PC61 group were less severe than those in control group. Conclusions Murine Tregs increase dramatically after Mycobacterium tuberculosis infection. Treg could inhibit the specific cellular immunity against Mycobacterium tuberculosis, and therefore, may facilitate the persistent infection of Mycobacterium tuberculosis and development of tuberculosis.

A high efficient in vitro regeneration protocol was developed from leaf explants of the female 'Red Sun' kiwifruit (Actinidia chinensis) and the multiplication coefficient and rooting rate of adventitious buds were also optimized. This method does not require formation of callus tissues which leads to somaclonal variations. The results show that the adventitious buds developing directly from explants tissue were noticed after 30 d of culture. The maximum regeneration frequency of adventitious buds is 100% and 18.67 shoots was observed in each leaf explants when MS medium was supplemented with 3.0 mg/L BA+1.0 mg/L NAA. The optimal culture medium for bud multiplication is MS+2.0 mg/L BA+1.0 mg/L NAA+0.1 mg/L GA3 and the multiplication coefficient reached 8.63. On the rooting medium with 1/2 MS+0.8 mg/L IBA for 15 d, the adventitious plantlets were transferred into matrix perlite supplied with 1/2 MS liquid medium for 15 d and the rooting rate reached 100%. 95 out of 98 plantlets (96.94%) survived acclimatization, producing healthy plants in the greenhouse. Taken together, a highly efficient regeneration method via leaf explants of 'Red Sun' kiwifruit was successfully established. This protocol may be useful for micropropagation and genetic transformation studies of 'Red Sun' kiwifruit.
Actinidia ; growth & development ; Plant Growth Regulators ; pharmacology ; Plant Leaves ; growth & development ; Regeneration ; physiology ; Tissue Culture Techniques ; methods

Bacterial canker caused by Pseudomonas syringae pv. Actinidiae is one of the most important diseases of kiwifruit (Actinidia chinensis) and leads to considerable yield losses. In order to obtain transgenic plants with resistance for 'Red Sun' kiwifruit to canker disease, a non-specific lipid transfer protein-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) was introduced into 'Red Sun' kiwifruit through Agrobacterium-mediated transformation. After two days of co-cultivation with A. tumefaciens strain LBA4404 harboring 35S:LJAMP2, the transformed explants were transferred to the selection medium containing 25 mg/L kanamycin+3.0 mg/L BA+1.0 mg/L NAA. The regeneration efficiency of kanamycin-resistant shoots reached to 85%. All (100%) of kanamycin-resistant shoots rooted on half-strength MS medium supplemented with 0.8 mg/L IBA and a total of 40 regenerated plantlets were obtained. PCR and histochemical GUS activity analysis show that 23 of 40 lines (57.50%) were positive, suggesting that the LJAMP2 gene was integrated into the genome of 'Red Sun' kiwifruit. Taken together, we established an efficient genetic transformation method for 'Red Sun' kiwifruit using A. tumefaciens and the transformation frequency reached 5.11%. This protocol will be useful for the genetic breeding of 'Red Sun' kiwifruit for improvement of disease resistance.
Actinidia ; genetics ; Agrobacterium ; Antigens, Plant ; genetics ; Carrier Proteins ; genetics ; Leonurus ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; Transformation, Genetic

OBJECTIVETo investigate the effect of SIRT6/NF-κB signal axis in delaying hematopoietic stem/progenitor cell senescence with ginsenoside Rg1, in order to provide theatrical and experimental basis for looking for methods for delaying HSC senescence.

METHODSca-1 + HSC/HPC was isolated by magnetic cell sorting (MACS) and divided into five groups: the normal control group, the aging group, the positive control group, the Rg1 anti-senescence group, and the Rg1-treated group. Senescence-associated β-galactosidase (SA-β-Gal) staining, cell cycle analysis and hemopoietic progenitor cell mix (CFU-Mix) were adopted to determine the effect Rg1 in delaying or treating Sca-1 + HSC/HPC senescence biology. The mRNA and protein of senescence regulation molecules SIRT6 and NF-KB were examined by realtime fluorescence quantitative PCR (FQ-PCR) and western blotting.

RESULTCompared with the senescence group, the Rg1 anti-senescence group and the Rg1-treated group showed lower percentage in SA-β-Gal-stained positive cells, decreased cell proportion in G1 phase, increased number of CFU-Mix, up-regulated in SIRT6 mRNA and protein expression, down-regulation in NF-KB mRNA and protein expression. The Rg1 anti-senescence group showed more evident changes in indexes than the Rg1-treated group.

CONCLUSIONRg, may inhibit Sca-1 + HSC/HPC senescence induced by t-BHP by regulating SIRT6/NF-KB signal path.

Animals ; Antigens, Ly ; analysis ; Cellular Senescence ; drug effects ; Female ; Ginsenosides ; pharmacology ; Hematopoietic Stem Cells ; drug effects ; Male ; Membrane Proteins ; analysis ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; physiology ; Signal Transduction ; physiology ; Sirtuins ; physiology