Objective To study on immunity function in infectious mononucleosis (IM) children infected by Epstein-Barr virus (EBV) and its relationship with clinic and prognosis.Methods Serum immunoglobulin was detected by immunoradiometric nephelometry,expressions of C D3+,CD4+,CD8+,CD19+,CD23+ on T-lymphocytes subset in peripheral blood were measured by flow cytometry in 50 IM (IM group)who was in acute and convalescent period,and compared with 50 healthy controls (control group).Results The levels of IgA,IgG,IgM in IM group with acute period were (0.75 ± 0.65),(7.55 ± 2.05),(1.85 ± 0.55)g/L,in IM group with convalescent period were (0.95 ± 0.55),(8.85 ± 2.25),(1.75 ± 0.65) g/L.In control group,those were (1.25 ± 0.75),(10.65 ± 2.55),(1.80 ± 0.50) g/L.IgA and IgG in IM group with acute period were significantly lower than those in IM group with convalescent period and control group (P ＜0.01),but IgM was no significant difference among them (P ＞0.05).The levels of CD3+,CD4+,CD8+,CD4+/CD8+,CD19+,CD23+ in IM group with acute period were 0.6050 ± 0.0850,0.2080 ± 0.0315,0.6520 ± 0.0520,0.45 ± 0.35,0.0580 ± 0.0205,0.0250 ± 0.0135,in IM group with convalescent period were 0.7220 ± 0.0820,0.3575 ± 0.0375,0.3565 ± 0.0565,1.45 ± 0.45,0.1580 ± 0.0280,0.0625 ± 0.0225.In control group,those were 0.7530 ± 0.0830、0.4850 ± 0.0450、0.3275 ± 0.0575 1.48 ± 0.55、0.1850 ± 0.0560、0.0805 ± 0.0175.CD3+,CD4+,CD4+/CD8+,CD19+,CD23+ in IM group with acute period were significantly lower than those in IM group with convalescent period and control group (P ＜ 0.05),but CD8+ was significantly higher than that in IM group with convalescent period and control group (P ＜0.05).Conclusion The abnormality of immunoglobulin and T-lymphocytes subset in children infected by EBV is obvious.

Arnebia euchroma is the main source for medicinal herb Zicao. and its most important component shikonin compounds have high medicinal and industrial value. This research is aimed to build overexpression vectors and RNAi vectors for key secondary metabolism genes of A. euchroma, and bulid platform for constructions of related transgenic lines using GATEWAY technology. To build genetic material based genetic research platform is to provide a great convenience for digging and functional verification of the genes on secondary metabolic pathway, and also to fill the gaps in transgenic research of A. euchroma. This study is also important for the cultivation of shikonin high-yielding strains of A. euchroma.
Boraginaceae ; genetics ; metabolism ; Genetic Vectors ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; RNA Interference ; Secondary Metabolism

Twenty-seven ERF transcription factor family genes were isolated from Arnebia euchroma, with an average size of 1,010 bp, each gene encoded a 212 amino acids on average. The gene structure and expression of physicochemical properties, subcellular localization, signal peptides, senior structural domains and conservative forecasting, and analysis of A. euchroma were studied comparing with ERF gene gi261363612 of Lithospermum erythrorhizon, and phylogenetic analysis of A. euchroma ERF family was carried out. The results showed the existence of three conserved domains in this family, the senior structure based on random coil and it clustered into CBF/DREB and ERF subfamilies.
Amino Acid Motifs ; Amino Acid Sequence ; Boraginaceae ; genetics ; Cloning, Molecular ; Computational Biology ; Genome, Plant ; genetics ; High-Throughput Nucleotide Sequencing ; Medicine, Chinese Traditional ; Molecular Sequence Data ; Multigene Family ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; Plants, Medicinal ; Protein Structure, Secondary ; Protein Structure, Tertiary ; Sequence Analysis, DNA ; Transcription Factors ; chemistry ; genetics ; Transcriptome

A cDNA sequence of Arnebia euchroma AP2/ERF named AeAP2/ERF was cloned by in silico cloning in this study, using ACX71873 sequence from Lithospermum erythrorhizon as the probe sequence. Some characters of the AP2/ERF gene and encoded protein sequences were predicted and analyzed by the bioinformatics methods, including general physical and chemical properties, hydrophobieity, signal peptide, secondary structure, localization sites in cells. Results showed that the 876 bp long gene included a 1 077 bp ORF and encoding 205 amino acid. The AeAP2/ERF protein had no signal peptide, it was a hydrophilic proteins located in nucleus. The function of the AP2/ERF protein was mainly involved with metabolism controlling and signal transduction.
Amino Acid Sequence ; Base Sequence ; Boraginaceae ; classification ; genetics ; Cloning, Molecular ; Computational Biology ; Computer Simulation ; Hydrophobic and Hydrophilic Interactions ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Protein Structure, Secondary ; Protein Structure, Tertiary ; Transcription Factors ; chemistry ; genetics

OBJECTIVETo explore the molecular mechanism of Wenxin Capsule (WXC, an effective Chinese composite drug) in preventing and treating myocardial ischemia of coronary heart disease.

METHODSRat model of myocardial ischemia was established by subcutaneous multi-point injecting isoproterenol. Effect of WXC on cell apoptosis was observed by transmission electron microscopy and TUNEL method, and its effect on apoptotic related gene Bcl-2 and Fas gene protein expression was observed by immunohistochemical method.

RESULTSIsoproterenol induced myocardial ischemic injury could cause evident cardial cell apoptosis, obvious enhance Fas gene protein expression and mild enhance Bcl-2 gene protein expression. WXC could significantly down-regulate Fas, up-regulate Bcl-2 gene protein expression, significantly inhibit and block the myocardial cell apoptosis.

CONCLUSIONSTo inhibit and block the event of cell apoptosis through regulating Bcl-2 and Fas gene protein expression in ischemic myocardium might be one of the mechanisms of WXT in preventing and treating myocardial ischemic injury of coronary heart disease.

Animals ; Apoptosis ; drug effects ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Isoproterenol ; Male ; Myocardial Ischemia ; chemically induced ; metabolism ; pathology ; Myocytes, Cardiac ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; fas Receptor ; biosynthesis ; genetics

Background C57BL/6(B6) is a kind of routine mouse specie used in experimental autoimmune uveitis (EAU) research.Previous studies showed that the pathogenesis of uveitis related to inflammatory cytokines secreted by different helper T(Th) cells.However,the interaction of different Th cells in EAU is unclear.Objective This study was to investigate the dynamic changes of inflammatory factors in the spleen and serum after immunization in EAU mice.Methods Forty-four SPF B6 mice were immunized by injection of interphotoreceptor retinoid-binding protein (IRBP) and complete Freund adjuvant (CFA) emulsion via caudal vein and footpad.Indirect ophthalmoscope was used to examine the eyes 3 times per week and the inflammatory response was scored based on Thurau's criteria.In the thirty day after injection,20 model eyes were extracted and the sections of eye tissue were prepared for histopathological examination.The spleens of model mice were enucleated before injection and 2,5,10,15,20,25,30 days after injection,and reverse transcriptase PCR (RT-PCR) was used to detect the contents of interleukin-17 (IL-17) mRNA,interferon-γ (IFN-γ) mRNA,tumor necrosis factor-α (TNF-α) mRNA and IL-10 mRNA,and the contents of IL-17,IFN-γ,TNF-α and IL-10 in model serum were assayed by ELISA in 24 model mice.The experimental protocol and use of the animals were approved by Ethic Committee for Care and Use of Laboratory Animals of Shandong University of Traditional Chinese Medicine.Results Mild inflammatory response was seen in 12 days under the indirect ophthalmoscope with the scores of 0.5.The inflammatory scores peaked in 13-15 days with the scores of 1.0 and alleviated after that with the inflammatory scores of 0.5 in 30 days after injection.The histopathological score was consistent with the clinical score in the models on the 30 days.The serum IL-17 content of model mice was (0.98±0.05) ng/L before injection and increased to (51.85 ±2.42) ng/L on the fifth day,and decreased to (4.01±0.06)ng/L on the fifteen day.But,the serum IL-17 level increased to (25.00±0.94)ng/L again on the 25th day,and then lowed to (6.01 ±0.21)ng/L 30 days after injection,showing a significant elevation in comparison with that of before injection (P=0.000).The serum IFN-γ content of the model mice was (1.02±0.09)ng/L before injection and increased to (50.54±0.48) ng/L on the fifth day,and (73.21±0.12) ng/L on the tenth day,and then it declined gradually until (5.15±0.18)ng/L in the 30th day,which was still higher than that of before injection (P=0.000).After injection of IRBP+CFA,the serum TNF-α level upregulated from the second day to fifth day with the peak values (134.25±0.59)ng/L,and declined to valley on 15th day.A repeat elevation of serum TNF-α level was found on the 20th day with the values (60.54±0.62)ng/L and followed by decrease till the 30th day,which was higher than that of before injection (P=0.660).Serum IL-10 was detectable in the tenth day and peaked on the 15th day.Then a slight decrease was seen till the 30th day,compared with before injection(P =0.000).The contents of IL-10 mRNA,IL-17 mRNA,TNF-α mRNA,IFN-γmRNA in mice spleens followed the same pattern with serum levels of their proteins.Conclusions IL-17,IFN-γ,TNF-α and IL-10 are key inflammatory factors of Th1,Th2 and Th17,they present with specific changes during EAU,it confirming that IFN-γ probably play a pathogenic role in EAU,IL-17 and TNF-α levels probably associated with the chronic and recurrent procedure of uveitis,IL-10 plays an inhibit role in EAU.

Objective To localize the tibial attachment of the posterior cruciate ligament (PCL) on the magnetic resonance imaging (MRI) and provide parameters for clinical PCL reconstruction.Methods We retrospectively analyzed 524 patients with intact tibial PCL attachment who had undergone knee MRI from January 2010 to January 2016.They were 286 men and 238 women with an average age of 35 years (from 20 to 50 years).The size and positions of the tibial PCL attachment were measured on the sagittal and coronal MRI slices.The differences were analyzed between different genders.Results On the sagittal slices,the mean distance from the central tibial PCL attachment to the posterior edge of the tibial plateau was 17.9 ± 3.0 mm and the mean anteroposterior diameter of the tibial PCL attachment was 9.7 ± 2.4 mm,with those for males significantly larger than for females (P ＜ 0.05).The above mean values when expressed as a percentage of the posterior tibial slop were 79.9％ ±4.5％ and 43.7％ ± 9.6％,respectively,showing no significant differences between males and females (P ＞ 0.05).On the coronal slices,the distances from the central tibial PCL attachment to the medial and lateral edges of the tibial plateau were 33.5 ± 3.1 mm and 37.4 ±4.1 mm,respectively,and the mediolateral diameter of the tibial PCL attachment was 12.0 ± 1.6 mm,with those for males significantly larger than for females (P ＜ 0.05).The above mean values when expressed as a percentage of the mediolateral diameter of the tibial PCL attachment were 47.4％ ± 3.2％,52.7％ ±3.1％ and 16.9％ ± 1.7％,respectively,showing no significant differences between males and females (P ＞ 0.05).Conclusions On knee MRI images,the distance from the central tibial PCL attachment to the posterior edge of the tibial plateau is about 17.9 mm,the anteroposterior diameter of the tibial PCL attachment around 9.7 mm,and the mediolateral diameter of the tibial PCL attachment roughly 12.0 mm.These measurements for males are larger than for females.

Objective To investigate the role of increased microRNA-21 (miR-21) in the development of renal tubulointerstitial fibrosis secondary to aristolochic acid induced acute kidney injury.Methods C57BL/6J male mice were intraperitoneally injected with aristolochic acid at a dose of 10 mg/kg.Blood samples and kidneys were harvested at day 1,3,7,14,28 after aristolochic acid treatment.To assess the role of miR-21 in aristolochic acid induced acute kidney injury to chronic kidney disease progression,mice were intravenously injected with anti-miR-21 or anti-scramble (10 mg/kg) at 1 h before aristolochic acid dosing,as well as d5 and d10 after aristolochic acid dosing.Results Increased serum creatinine and severe kidney injury were found at d3 after aristolochic acid treatment.Renal tubulointerstitial fibrosis was developed at d14 after aristolochic acid treatment.Protein expression of α-SMA,vimentin and collagen Ⅰ were significantly up-regulated at d7 and peaked at d14 (P ＜ 0.01),while protein abundance of E-Cadherin decreased at d14 and lasted until d28 (P ＜ 0.01).The abundance of miR-21 increased at d7 after aristolochic acid dosing,peaking at d14 and thereafter maintaining at a high level.Anti-miR-21 intervention relieved renal injury with reduced serum creatinine (P ＜ 0.05) and attenuation of renal tubulointerstitial fibrosis.Besides,the protein expression of α-SMA,vimentin,and collagen Ⅰ/Ⅳ was all down-regulated after anti-miR-21 treatment (P ＜ 0.05).PTEN was up-regulated and the ratio of its downstream genes p-AKT/AKT was decreased.(P ＜ 0.05) Conclusions A single high dose of aristolochic acid leads to acute kidney injury and the development of renal tubulointerstitial fibrosis secondary to AKI.Renal tubulointerstitial fibrosis could be partially reversed by inhibiting miR-21 via PTEN/p-AKT pathway.

Objective To study the effects of abiotic elicitors methyl jasmonate (MeJA) and salicylic acid (SA) on the alkaloids accumulation and related enzymes metabolism inPinellia ternata suspension cell cultures. Methods Using the leaf petioles-derived suspension cell cultures as the study object, the culture duration, concentrations of MeJA and SA were determined to get the optimal alkaloids accumulation, and the activities of metabolic enzymes IMP dehydrogenase and sAMP synthase were also measured.Results A 9-fold of dried biomass and a 3-fold of alkaloids accumulation were observed inP. ternata suspension cell cultures after culture for 21 d. Both MeJA and SA could significantly promote the accumulation of alkaloids inP. ternata suspension cells. 150 μmol/L MeJA enhanced alkaloids content (4.7 mg/gDW) by 3.6 folds in comparison with control group, whereas 50 μmol/L SA showed a 2.5-fold increase. Meanwhile, 100 μmol/L MeJA and 50 μmol/L SA promoted the increase in IMP dehydrogenase activity by 3.0 and 3.7 fold respectively, and 150 μmol/L MeJA and 100 μmol/L SA showed the increase by 2.6 and 4.4 fold respectively.Conclusion Proper adding exogenous MeJA and SA can promote the accumulation of alkaloids inPinellia ternata suspension cell cultures.

Objective:To investigate the safety and efficacy of laminectomy combined with vertebroplasty in spinal metastases from rapid-growth tumors. Methods:Clinical data of 23 patients with spinal metastases of lung cancer, who were admitted to the Cancer Hospital from July 2008 to May 2012, were retrospectively analyzed. Thirteen male and ten female patients, with an age range from 40 years to 65 years and a mean age of 51.5, were examined. All patients received posterior laminectomy to relieve spinal cord compression. Afterward, vertebroplasty combined with radiofrequency ablation was conducted, followed by the internal fixation of vertebrae (instrumental fixation). Operation time, blood loss, and bone cement leakage rate were analyzed. One month before and after the operation, pain measurement was conducted using visual analog scale (VAS) and neurologic deficit (spinal cord injury) by Frankel Grade. Functional impairment was classified by Karnofsky performance status (KPS) score. Quality of life was assessed by the European Organization for Research and Treatment questionnaire (EORTC QLQ-C30). Results:The mean operation time was 163±87.36 min. Blood boss was 430±130.35 mL. Bone cement leakage rate was 21.7%. One month before and after surgery, the VAS showed statistical significance (t=25.6, P<0.01). After surgery, 78.3%of all patients exhibited functionally satisfactory Frankel Grade D or E, compared with 43.5%of patients before the operation. KPS score (80 to 100) percentage was 69.6%after surgery compared with 34.8%before surgery. One month after the operation, remission of various degrees was seen in 10 of 18 patients who had sphincteric dysfunction before surgery (55.6%). The EORTC QLQ-C30 score was 85.39±8.99 before and 52.78±15.17 after operation. The quality of life improved significantly (t=11.6, P<0.01). Conclusion:Posterior laminectomy and vertebroplasty combined with radiofrequency ablation for spinal metastases from lung cancer is safe and effective. The treatment can improve pain, function, and life quality of patients with lung cancer spinal metastases.