In this paper, the chloroplast psbK-psbI intergenic spacers of 18 species of Dendrobium and their adulterants were amplified and sequenced, and then the sequence characteristics were analyzed. The sequence lengths of chloroplast psbK-psbI regions of Dendrobium ranged from 474 to 513 bp and the GC contents were 25.4%-27.6%. The variable sites were 71 while the informative sites were 46. The inter-specific genetic distances calculated by Kimura 2-parameter (K2P) of Dendrobium were 0.006 1-0.058 1, with an average of 0.028 4. The K2P genetic distances between Dendrobium species and Bulbophyllum odoratissimum were 0.093 2-0.120 4. The NJ tree showed that the Dendrobium species can be easily differentiated from each other and 6 samples of the inspected Dendrobium species were identified successfully through sequencing the psbK-psbI intergenic spacer. Therefore, the chloroplast psbK-psbI intergenic spacer can be used as a candidate marker to identify Dendrobium species and its adulterants.
Chloroplasts ; DNA, Chloroplast ; genetics ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; classification ; genetics ; Plants, Medicinal ; classification ; genetics

OBJECTIVETo investigate the pathogenesis of globozoospermia, fertilization ability of round-headed sperm, and the application value of assisted oocyte activation in intracytoplasmic sperm injection (ICSI) for the wives of glohozoospermia men.

METHODSWe collected oocytes from the wives of 2 globozoospermia patients and randomly divided them into two groups after ICSI to receive calcium ionophore A23187-activation and conventional treatment, respectively. We reviewed the relevant literature published at home and abroad, and discussed the etiology of globozoospermia, fertilization ability of round-headed sperm, and treatment options for this disease.

RESULTSQuality embryos were obtained in the A23187-activation group while no fertilized oocytes, oocyte cleavage, quality embryos, or blastular formation were found in the conventional treatment group. Both women achieved pregnancy and gave birth to healthy neonates after transfer of the quality embryos from the A23187-activation group.

CONCLUSIONCalcium ionophore A23187 can be applied to ICSI for the wives of globozoospermia men and bring about desirable clinical outcomes. Meanwhile, attention should be paid to its safety.

Calcimycin ; therapeutic use ; Calcium Ionophores ; therapeutic use ; Female ; Humans ; Infertility, Male ; drug therapy ; Male ; Oocytes ; Pregnancy ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; abnormalities

OBJECTIVETo observe the effectiveness and safety of external applying Compound Tripterygium wilfordii Hook F. (TwHF) in relieving joint pain in rheumatoid arthritis (RA) patients.

METHODSIn this double-blinded, randomized multicenter trial, a total of 174 moderately active RA patients were enrolled and randomly assigned to the treatment group (treated with Compound TwHF, 87 cases) and the placebo control group (87 cases). Compound TwHF or placebo was externally applied in painful joints, 20 g each time, once per day for 8 weeks. Self-reported joint pain relief was taken as a primary effective indicator. Visual analogue scale for pain (VAS), disease activity score of 28 joints (DAS28), VAS for general health (GH) were evaluated before treatment, at week 4 and after treatment. Erythrocyte sedimentation rate (ESR) and hypersensitive C reactive protein (hs-CRP) were tested before and after treatment. Menstrual changes in females were observed during treatment. Skin irritation occurred during the recording process was assessed using skin irritation strength. Intention to treat (ITT) was statistically analyzed.

RESULTSThe joint pain relief rate in the treatment group was 90.8% (79/87 cases), higher than that in the placebo control group (69.0%, 60/87 cases; P = 0.001). VAS pain score, DAS28, VAS for GH score were significantly improved in the two groups at week 4 of treatment and after treatment, as compared with before treatment (P < 0.01). ESR and hs-CRP levels significantly decreased in the treatment group after treatment (P < 0.05, P < 0.01). No difference was found in post-treatment VAS pain score, DAS28, VAS for GH score, ESR, or hs-CRP between the two groups (P > 0.05). Eight adverse events occurred in the treatment group (5 skin allergy, 1 intolerance of medical odor, and 2 mild liver injury), while 3 adverse events occurred in the placebo control group (2 skin allergy, 1 mild liver injury). There was no statistical difference in adverse event between the two groups (P > 0.05). No menstrual change occurred in the treatment group.

CONCLUSIONExternal applying Compound TwHF was an effective and safe way to relieve-joint pain of RA patients, which could be taken as an adjuvant therapy.

Arthralgia ; drug therapy ; Arthritis, Rheumatoid ; drug therapy ; Blood Sedimentation ; C-Reactive Protein ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Phytotherapy ; Treatment Outcome ; Tripterygium

BACKGROUNDPrompt diagnosis of Mycobacterium tuberculosis (MTB) infection is an essential step in tuberculosis control and elimination. However, it is often difficult to accurately diagnose pediatric tuberculosis (TB). The tuberculin test (TST) may have a low specificity because of cross-reactivity with antigens present in Mycobacterium bovis bacillus Calmette-Guerin (BCG) and other mycobacteria, especially in China with a predominantly BCG-vaccinated population. Early-secreted antigenic target 6-kDa protein (ESAT-6) and culture filtrate protein 10 (CFP-10), stand out as suitable antigens that induce an interferon-gamma (IFN-γ) secreting, T-cell-mediated immune response to infection. While, considered the higher costs and complexity of the IFN-γ release assay (TSPOT), we aimed to evaluate the TSPOT and TST test in the clinical diagnosis of pediatric tuberculosis and to establish a diagnostic process suitable for China.

METHODSThe sensitivity and specificity of the assay were evaluated in total seventy four children with active tuberculosis and fifty one nontuberculous children with other disease, and then the results were compared with TST. Logistic regression models were used to identify variables that were associated with positive results for each assay. The independent variables included sex, age, birth place, vaccination history, close contract with an active TB patient.

RESULTSThe sensitivity of TSPOT was higher than TST in active TB children with or without BCG vaccination, as well as in children with culture-confirmed TB. But the difference was not significant statistically. Combining results of the TSPOT and TST improved the sensitivity to 94.6%. Agreement of the TST and TSPOT was low (77.0%, κ = 0.203) in active TB patients. The difference in specificity between TSPOT and TST test was statistically significant (94.1% vs. 70.6%, P = 0.006). Specificity of the two tests in patients without prior BCG vaccination history was similar (80.0% vs. 60.0%). The concordance between the two tests results in BCG vaccinated subjects was low (71.7%, κ = 0.063). For TSPOT, none of the included risk factors was significantly associated with positive results. For TST, BCG vaccination (OR: 1.78; 95%CI: 1.30 - 2.00) was significantly associated with positive results.

CONCLUSIONSAlthough IFN-γ release assay had relatively high sensitivity and specificity, we also should consider the higher costs and complexity of this test. Therefore, TSPOT could be used as the complementary tool of TST in circumstances when a suspected patient with negative TST results, or to exclude a positive TST result caused by BCG vaccination.

BCG Vaccine ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Interferon-gamma ; secretion ; Logistic Models ; Male ; Sensitivity and Specificity ; Tuberculin Test ; methods ; Tuberculosis ; diagnosis ; Vaccination

AIM To prepare Scutellariae Radix total flavonoids-loaded liposomes and to evaluate their in vitro anti-tumor activity.METHODS Liposomes were prepared by reverse evaporating method.Taking phospholipid concentration,ratio of phospholipid to cholesterol,ratio of organic phase to aqueous phase and total flavonoids concentration as influencing factors,together with encapsulation efficiency as an evaluation index,Box-Behnken design was applied to optimizing the preparation,after which MTT was employed to detect the obtained liposomes' inhibitory effect on human hepatocellular carcinoma cells HepG2.RESULTS The optimal conditions were determined to be 30 mg/mL for phospholipid concentration,3 ∶ 1 for ratio of phospholipid to cholesterol,2 ∶ 1 for ratio of organic phase to aqueous phase,and 5 mg/mL for total flavonoids concentration.The obtained round and uniform liposomes demonstrated an average particle size of (160.7 ± 12.0) nm,Zeta potential of (-41.4 ±2.3) mV,encapsulation efficiency of (86.19 ±0.44)％,drug loading of (5.32 ± 0.04)％,and accumulative release rate of (80.77 ± 2.53) ％ at 24 h.Compared with total flavonoids,liposomes exhibited stronger inhibitory effect on HepG2 cells with the IC50 of 48.853 μg/mL at 48 h in a time-and dose-dependent manner.CONCLUSION Scutellariae Radix total flavonoids-loaded liposomes stored at low temperature (4 ℃) display sustained-release effect and in vitro anti-tumor activity.

The aim of present study was to investigate the effect of chronic intermittent hypobaric hypoxia (CIHH) on collagen-induced arthritis (CIA) in rat. Fifty male adult Sprague-Dawley rats were randomly divided into 5 groups: CIHH pre-treatment group (Pre-T), pre-control group (Pre-C), CIHH post-treatment group (Post-T), post-control group (Post-C) and blank control group (Con). The rats in Pre-T and Post-T groups were exposed to 28 d of hypobaric hypoxia (simulated 3 000 m altitude, 5 h per day, pO2 = 108.8 mmHg, 14% O2) in a hypobaric chamber before and 12 days after CIA induction, respectively. The rats in Pre-C and Post-C groups were only experienced CIA induction, being control groups for Pre-T and Post-T groups, respectively. The rats in Con group were not given any treatment. The thickness of two-hind paw of rat was measured with spiral micrometer and the degree of arthritis was evaluated by arthritis index (AI). Morphological changes of ankle joint were observed through HE staining. The apoptotic rate in synovial tissue was measured by terminal dUTP nick end labeling (TUNEL) and the apoptotic rate of CD3(+) T lymphocyte in spleen was measured by flow cytometry technique. The protein expressions of Bcl-2 and Bax were measured using immunohistochemistry SP method. The results showed that incidence rate of CIA in Pre-T rats was lower than that in Pre-C rats (P < 0.05). AI in Pre-T and Post-T rats were smaller than those in Pre-C and Post-C, respectively (P < 0.05). In Pre-C and Post-C rats, there were hyperplasia of synovial cell, pannus forming, infiltration with inflammatory cell, and destroyed cartilage and bone in ankle joint. On the contrary, pathological changes of ankle joint were alleviated significantly in Pre-T and Post-T rats. Compared with Pre-C and Post-C rats, apoptotic rates of synovial cell and T lymphocyte in Pre-T and Post-T rats were increased (P < 0.05). As to the possible anti-apoptosis mechanism, CIHH, no matter before and after CIA induction, decreased Bcl-2 expression and increased Bax expression in joint synovial cells and spleen T lymphocytes (P < 0.05), respectively. In conclusion, CIHH possesses a protective effect against CIA in rat by enhancing apoptosis of synovial cells and T lymphocytes, which may be related to the inhibition of Bcl-2 protein expression and the promotion of Bax protein expression.
Animals ; Apoptosis ; physiology ; Arthritis, Experimental ; pathology ; physiopathology ; prevention & control ; Hypoxia ; physiopathology ; Ischemic Postconditioning ; methods ; Ischemic Preconditioning ; methods ; Joints ; pathology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology ; Synovial Membrane ; pathology ; T-Lymphocytes ; pathology ; bcl-2-Associated X Protein

Objective: To analyze the clinical characteristics and long-term outcomes with multicenter study for acute lymphoblastic leukemia (ALL) in children over 10 years old and adolescents. Method: Newly diagnosed ALL patients aged from 10 to 18 years old in three hospitals were included in the study from May 1(st) 2005 to April 30(th) 2015. They were received ALL-2005/2009 protocol following up to December 31(st) 2016. The clinical characteristics, outcomes and the prognostic analysis were evaluated between the two protocols. Results: Totally, 237 patients were involved in the study, 76 cases for ALL-2005 and 161 cases for ALL-2009 protocol. Complete remission (CR) after induction therapy was 94.5%. 64 (28.6%) patients relapsed with a median time of 14.5 months and 70 (29.5%) patients passed away during the following time. In long-term follow-up, the 5-year event-free survival (EFS) and 5-year overall survival (OS) of ALL patients were (63.1±3.3)% and (68.4±3.2)%. The 7-year EFS and OS were (61.0±3.5)% and (67.6±3.3)%.The 5-year EFS of intermediate risk group in ALL-2005 and ALL-2009 protocol were (73.6±6.1)% and (71.7±4.3)% with no difference (χ(2)=0.064, P=0.801). The 5-year EFS of high risk group in two protocols were (27.6±9.6)% and (33.9±9.3)%, showing no significant difference (χ(2)=0.296, P=0.586). Five years relapsed rate of two protocols were (33.8±5.7)% and (32.6±4.1)% with no difference (χ(2)=0.055, P=0.815). The mortalities were 36.8% and 29.8% separately (χ(2)=2.869, P=0.090). Univariate analysis indicated that age, male, risk, BCR/ABL translocation/t(9;22) and resistant to induction were risk prognostic factors in long-term survival (χ(2)=4.764, 4.796, 46.410, 9.560, 25.450; P=0.029, 0.029, <0.001, 0.049, <0.001). Cox multivariate analysis showed male, risk and resistant to induction were independent risk prognostic factors (RR=1.790, 2.727, 2.719; P=0.021, 0.000, 0.012). Conclusion: Protocol ALL-2009 enhanced the chemotherapy intensity in intermediate risk group with no benefit of survival. BCR-ABL fusion or t(9;22) translocation was still the risk factor of prognosis. TKI inhibitor used in these patients could improve survival. EFS rate was increased a little and death rate was decreased in ALL-2009 protocol with no significant lower relapsed rate comparing with ALL-2005 protocol.
Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; Child ; Disease-Free Survival ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Prognosis ; Remission Induction ; Risk Factors ; Translocation, Genetic

OBJECTIVETo evaluate the colouration of zirconia ceramic by adding three kinds of rare earth oxides. The influence of the pigments concentration on the mechanical properties and the microstructure was also analyzed.

METHODSAdded different concentrations of CeO(2), Er(2)O(3) and Pr(6)O(11) in tetragonal zirconia poly crystals stabilized with 3 mol% yttria (3Y-T2P) powder, compacted at 200 MPa using cold isostatic pressure, and sintered to 1 400 degrees C. The heating rate was 150 degrees C/h and the dwelling time was 2 hours. The chromaticity of sintered bodies was measured with chroma meter. The relative density, hardness, flexure strength and fracture toughness were investigated as well. The phase stability of the colorized and pure zirconia was evaluated by X-ray diffraction (XRD) using an automated diffractometer. The microstructures of the specimens were evaluated by scanning electron microscope (SEM).

RESULTSSeveral kinds of color achieved by the different pigments praseodym oxide, cerium oxide and erbium oxide were presented in the CIELab system. The a* value increased with the added amount of Er(2)O(3), while b* value rose with the increasing amount of CeO(2) and Pr(6)O(11). However, three pigments failed to decrease L* value and the sintered body appeared too bright. Adding three pigments influenced flexure strength of zirconia ceramic significantly, but had little influence on the hardness and fracture toughness. Microscopy revealed the relationship between the porosity and shapes of grains was correlated to strength of the diphase ceramics. No additional phase could be detected by XRD, except t-ZrO(2) in all colorized samples after sintering at 1 400 degrees C for 120 min.

CONCLUSIONSZirconia ceramic can be colorized by CeO(2), Er(2)O(3), and Pr(6)O(11). Pigments even in a small amount influence the mechanical properties of the colorized zirconia material, which necessitates further investigation.

Coloring Agents ; pharmacology ; Dental Porcelain ; chemistry ; Dental Stress Analysis ; Metals, Rare Earth ; pharmacology ; Oxides ; pharmacology ; Prosthesis Coloring ; Zirconium ; chemistry

OBJECTIVETo construct one recombinant vaccinia virus expressing the HPV18 E6 and E7 fusion proteins as HPV18 therapeutic vaccine candidate, and test its immunogenicity.

METHODSThe fusion E7E6 genes were synthesized and mutated to inactivate their oncogenic potential, and inserted into a vaccinia virus plasmid vector to construct one recombinant vaccinia virus. Finally its immunogenicity was characterized in immunized mice.

RESULTSOne recombinant vaccinia virus expressing HPV18 E7E6 fusion proteins was constructed. Sequencing results of PCR products and Western blot tests showed that the E7E6 fusion genes were correct and expressed in CEF cells infected with the recombinant vaccinia virus. The specific antibodies against E6 and E7 proteins were elicited, however no positive responses were detected by ELISPOT in immunized mice.

CONCLUSIONSOne recombinant vaccinia virus expressing HPV18 E7E6 fusion proteins was generated and elicited specific antibodies against E6 and E7 proteins, but detected no positive cellular immune responses in immunized mice, which will provide the basis to develop the different animal model for examining the cellular immune responses of HPV18E6 and E7 proteins.

Animals ; Antibodies, Viral ; blood ; DNA-Binding Proteins ; genetics ; immunology ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Human papillomavirus 18 ; genetics ; immunology ; Humans ; Immunization ; Male ; Mice ; Mice, Inbred C57BL ; Oncogene Proteins, Viral ; genetics ; immunology ; Papillomavirus Infections ; immunology ; virology ; Papillomavirus Vaccines ; genetics ; immunology ; Random Allocation ; Recombinant Fusion Proteins ; genetics ; immunology ; Vaccinia virus ; genetics ; metabolism

OBJECTIVETo investigate the relationship between EGFR activation and down-regulation of miRNA-145 in lung cancer.

METHODSNormal human lung epithelia cell line (BEAS-2B), human lung adenocarcinoma cell lines with wild-type EGFR (A549 and H292) and human lung adenocarcinoma cell lines with EGFR mutation (H1975 and H1650) were chosen in this study. The levels of miRNA-145 and p-EGFR were determined by quantitative real-time PCR (qRT-PCR) and Western blotting, respectively, and the relationship between p-EGFR and miRNA-145 levels was analyzed. The miRNA-145 levels were determined by qRT-PCR after activating EGFR with EGF or blocking EGFR signal pathway with AG1478. In addition, ERK1/2 inhibitor U0126 was used to inhibit ERK1/2 activation and then the expression of miRNA-145 was detected.

RESULTSThe miRNA-145 levels were closely negatively related with p-EGFR in lung cancer cells (r = -0.926, P = 0.024). EGF down-regulated miRNA-145 expression, particularly in BEAS-2B cells (53.0%; t = 30.993, P = 0.001) and A549 cells (42.6%; t = 14.326, P = 0.005).The miRNA-145 was up-regulated after inhibiting p-EGFR with AG1478, and significantly enhanced by 67.5% in H1975 cells when treated with AG1478 (t = 8.269, P = 0.014). The ERK1/2 signal pathway was activated by p-EGFR. U0126 restored the miRNA-145 down-regulation induced by EGFR-activation in lung cancer cells.

CONCLUSIONThe activation of EGFR down-regulates miRNA-145 expression through ERK1/2 in lung cancer cells.

Butadienes ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Line ; Cell Line, Tumor ; Down-Regulation ; Enzyme Activation ; drug effects ; Enzyme Inhibitors ; pharmacology ; Epidermal Growth Factor ; pharmacology ; Epithelial Cells ; metabolism ; Humans ; Lung ; cytology ; Lung Neoplasms ; metabolism ; pathology ; MAP Kinase Signaling System ; MicroRNAs ; metabolism ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Nitriles ; pharmacology ; Phosphorylation ; Quinazolines ; pharmacology ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; metabolism ; Tyrphostins ; pharmacology