OBJECTIVE: To prepare paclitaxel loaded D-α-tocopheryl polyethylene glycol 1000 succinate(TPGS)-modified carboxymethyl chitosan-rhein polymeric micelles (PTX/TPGS-CR PMs) and preliminarily evaluate their performance. METHODS: PTX/TPGS-CR PMs was prepared by dialysis method, and the preparation procedure of PTX/TPGS-CR PMs was optimized by single factor with the drug loading, encapsulation rate and particle size as the indicators, then the optimized preparation procedure was verified. The safety of PTX/TPGS-CR PMs was initially investigated by the hemolysis test and the vascular irritation test. The cytotoxicity of PTX/TPGS-CR PMs in Hela cells was studied by MTT assay. Cell uptake experiments were performed by laser confocal microscopy and flow cytometry to investigate the uptake of PTX/TPGS-CR PMs by Hela cells. RESULTS: The particle size and PDI of PTX/TPGS-CR PMs prepared by the optimized preparation were (197.3±4.4) nm and (0.131±0.021), respectively. The Zeta potential was (-31.8±0.5) mV. The drug loading and encapsulation efficiency were (48.20±3.03)% and (87.26±4.91)%, respectively. The hemolytic test results showed that the hemolysis rate was less than 1.71%. No obvious irritation was observed after intravenous injection. The cytotoxicity of PTX/TPGS-CR PMs in Hela cells was concentration-and time-dependent. Cell uptake experiments showed that PTX/TPGS-CR PMs could be efficiently uptake by Hela cells. CONCLUSION: The PTX/TPGS-CR PMs has high drug loading and encapsulation efficiency, good safety. And they exhibite slightly better antitumor activity in vitro than Taxol®.

Objective:To investigate the effect of electroacupuncture (EA) at Ganshu (BL 18) and Shenshu (BL 23) on vascular endothelial growth factor (VEGF) and platelet endothelial cell adhesion molecule-1 (PECAM-1)/CD31 around the cerebral infarction focus in middle cerebral artery occlusion (MCAO) rats and the possible mechanism, thus to provide a new strategy for the treatment of cerebral ischemic stroke by acupuncture. Methods:A total of 180 healthy male Sprague-Dawley (SD) rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, 45 rats in each group. MCAO model was established using the modified line-embolus method in all rats except for those in the sham operation group; rats in the acupoint group were treated with EA at Ganshu (BL 18) and Shenshu (BL 23); rats in the non-acupoint group were treated with EA at the control points; rats in other 2 groups were only subjected to bundling without treatment. Ten rats in each group were randomly selected on the 3rd day, the 14th day and the 21st day after acupuncture stimulation to test the neurological function impairment. The expression levels of CD31 and VEGF were also detected. Results:Compared with the model group and non-acupoint group, the neurological function score of the acupoint group was decreased at each time point, and the differences were statistically significant (P<0.05,P<0.01). The expressions of VEGF and CD31 in each group were the lowest on the 3rd day, reached the peak on the 14th day and still remained at high level on the 21st day. And the differences among groups were statistically significant both on the 14th day and the 21st day (P<0.05,P<0.01). Compared with the model group and the non-acupoint group, the expressions of VEGF and CD31 in the acupoint group were increased, and the differences were statistically significant (allP<0.05). Conclusion: EA at Ganshu (BL 18) and Shenshu (BL 23) can significantly improve the neurological function score of MCAO model rats, and shows protective effect on cerebral ischemia. The protective mechanism may be related to the up-regulation of CD31 and VEGF expression around the cerebral infarction focus in the MCAO model rats and induction of angiogenesis.

To explore the association of serum Dickkopf-1 (DKK-1) levels with the development of ankylosing spondylitis (AS) and rheumatic arthritis (RA) in humans, databases including PubMed, EBSCO, Springerlink, Ovid, WANFANG and China National Knowledge Infrastructure (CNKI) were searched to identify relevant studies. On the basis of rigorous inclusion and exclusion criteria, case–control studies of the relationships between serum DKK-1 levels and AS and RA published before December 2014 were enrolled. Statistical analyses were performed using Comprehensive Meta-analysis 2.0 (CMA 2.0). Seven case–control trials with a total of 300 AS patients, 136 RA patients and 232 healthy controls were included in this study. Meta-analysis results revealed that DKK-1 serum levels were significantly higher in AS patients than in normal controls (standard mean differences (s.m.d.)=0.301, 95% confidence interval (CI)=0.094–0.507, P=0.004), whereas no significant difference in DKK-1 serum levels was observed between RA patients and healthy controls (s.m.d.=0.798, 95% CI=−2.166–3.763, P=0.598). Serum DKK-1 level may be closely related to the development of AS but not of RA.
China ; Humans ; Rheumatic Fever* ; Spondylitis, Ankylosing*

To explore the association of serum Dickkopf-1 (DKK-1) levels with the development of ankylosing spondylitis (AS) and rheumatic arthritis (RA) in humans, databases including PubMed, EBSCO, Springerlink, Ovid, WANFANG and China National Knowledge Infrastructure (CNKI) were searched to identify relevant studies. On the basis of rigorous inclusion and exclusion criteria, case–control studies of the relationships between serum DKK-1 levels and AS and RA published before December 2014 were enrolled. Statistical analyses were performed using Comprehensive Meta-analysis 2.0 (CMA 2.0). Seven case–control trials with a total of 300 AS patients, 136 RA patients and 232 healthy controls were included in this study. Meta-analysis results revealed that DKK-1 serum levels were significantly higher in AS patients than in normal controls (standard mean differences (s.m.d.)=0.301, 95% confidence interval (CI)=0.094–0.507, P=0.004), whereas no significant difference in DKK-1 serum levels was observed between RA patients and healthy controls (s.m.d.=0.798, 95% CI=−2.166–3.763, P=0.598). Serum DKK-1 level may be closely related to the development of AS but not of RA.
China ; Humans ; Rheumatic Fever* ; Spondylitis, Ankylosing*

OBJECTIVETo identify the candidate genes within the putative susceptibility locus for systemic lupus erythematosus (SLE) at 12p12.3-13.2.

METHODSKLRC1 was selected as the candidate gene according to the results of previous gene chip studies. TaqMan real-time quantitative PCR was performed for detecting KLRC1 mRNA expression in 55 SLE patients and 30 controls.

RESULTS AND CONCLUSIONKLRC1 mRNA expression was significantly higher in the mononuclear cells and T cells of SLE patients than in the healthy controls (P<0.01), but showed no significant difference in the B cells. No obvious correlation was found between the SLE disease activity index (SLEDAI) and KLRC1expression level, suggesting that KLRC1 can be a probable candidate gene for SLE on 12p12.3-13.2, but which is not associated with the disease activity.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; China ; Chromosomes, Human, Pair 12 ; genetics ; Female ; Gene Expression Profiling ; Genetic Predisposition to Disease ; Humans ; Lupus Erythematosus, Systemic ; ethnology ; genetics ; pathology ; Male ; Middle Aged ; NK Cell Lectin-Like Receptor Subfamily C ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Severity of Illness Index ; Young Adult

BACKGROUND: Proximal femoral nail anti-rotation is widely used to treat various intertrochanteric fractures. Although its operation trauma is small, and the blood loss of perioperative period is still large. Tranexamic acid has been gradually used to reduce the bleeding of intertrochanteric fracture. The effectiveness and safety of reducing blood loss during perioperative period were not reported. OBJECTIVE: To explore the safety and efficacy of tranexamic acid on perioperative blood loss in patients with intertrochanteric fracture undergoing proximal femoral nail anti-rotation. METHODS: One hundred and eight patients with intertrochanteric fracture undergoing proximal femoral nail anti-rotation were selected from First Affiliated Hospital, Guangzhou University of Chinese Medicine between January 2015 and January 2017. Among all the subjects, 52 patients who received the operation before January 2016 served as the control group and 56 patients who received the operation after January 2016 were selected as the treatment group. Half an hour before operation, patients in the treatment group received 1 g tranexamic acid dissolved in 250 mL normal saline by intravenous dropping; patients in the control group just received 250 mL normal saline by intravenous dropping. The bleeding volume, blood transfusion volume, hemoglobin, hematocrit, coagulation index, D-dimer levels and complications were compared between the two groups. RESULTS AND CONCLUSION: (1) During perioperative period, actual blood loss, intraoperative blood loss, dominant blood loss, recessive blood loss, volume of drainage, blood transfusion volume and blood transfusion rate were lower in the treatment group than in the control group (P < 0.05). (2) There was no statistically significant difference in the hemoglobin and hematocrit between the two groups before operation (P > 0.05). The hemoglobin and hematocrit of the two groups gradually decreased after the operation, and there was a slight improvement in the fifth day after surgery. At postoperative 2 hours, 1, 3 and 5 days, the hemoglobin and hematocrit of the treatment group were higher than in the control group (P < 0.05). At preoperation and each time point postoperation, prothrombin time, activated partial thromboplastin time, and fibrinogen levels were not statistically significant between the two groups (P > 0.05). Postoperative D-dimer levels in the two groups were significantly higher than preoperation, and there was a return on the fifth day. There was no statistically significant difference between groups at preoperation and each time point of postoperation (P > 0.05). (3) The results suggest that the tranexamic acid can effectively reduce the dominant and recessive blood loss in patients with the intertrochanteric fracture, and it is safe and effective.

OBJECTIVETo investigate the effect of sumoylation of alpha-synuclein by SUMO-1 on the mitochondria subcellular localization of alpha-synuclein and its degradation via ubiquitin-proteasome system.

METHODSPrimers of wild-type, A53T pathogenic mutant and K96R mutant of human alpha-synuclein were designed to amplify the corresponding cDNAs without stop codon. The cDNAs were cloned into pGEM T-easy vector, analyzed by using enzyme mapping and DNA sequencing, and subcloned into pEGFP-N1 vector. The recombinant plasmids of pEGFP-alpha-synuclein-WT, pEGFP-alpha-synuclein-A53T and pEGFP-alpha-synuclein-K96R were transfected into HEK293 cells by lipofectamine method. The expression of the alpha-synuclein protein was measured by immunofluorescence and confocal microscope. Then mitochondria staining as well as immunofluorescence were utilized to investigate the effect of wild-type, A53T mutant and sumoylation of alpha-synuclein on mitochondria subcellular localization of alpha-synuclein. The effect of sumoylation of alpha-synuclein on its degradation via the ubiquitin-proteasome system in the cells was assayed by Western-blot.

RESULTSThe enzyme mapping suggested that the eukaryotic expression plasmids for human wild-type, A53T and K96R mutants of the alpha-synuclein gene were constructed successfully. By immunofluorescence and confocal microscope, it was observed that alpha-synuclein-WT and alpha-synuclein-A53T proteins aggregated in cytoplasm, and alpha-synuclein-K96R protein aggregation was decreased in cytoplasm of cultured cells. The alpha-synuclein proteins of wild-type, A53T and K96R mutants were co-localized with mitochondria. Western-blot analysis revealed that both wild-type and A53T mutant affected the amount of the ubiquitinated proteins.

CONCLUSIONNeither overexpression of wild-type and A53T pathogenic mutant alpha-synuclein, nor sumoylation of alpha-synuclein, affected the subcellular localization in the mitochondria. However, overexpression of wild-type and A53T mutant alpha-synuclein affected the amount of the ubiquitinated proteins.

Blotting, Western ; Cell Line ; Humans ; Mitochondria ; metabolism ; Proteasome Endopeptidase Complex ; metabolism ; SUMO-1 Protein ; metabolism ; Ubiquitin ; metabolism ; alpha-Synuclein ; metabolism

OBJECTIVETo investigate the expression and role of glucose transporter-1 (Glut1) in infantile hemangioma.

METHODSFifty-two samples from infantile hemangioma, 25 in cavernous venous malformation, 9 in arteriovenous malformation, 2 in capillary malformation and 5 in normal skin samples were involved in this study. The EnVision immunohistochemical stain was used to investigate the expression of Glut1 protein in these samples.

RESULTSIn the early proliferating stage, a number of endothelial cells expressed Glut1. In the middle proliferating stage, most of vascular endothelial cells and scattered endothelial cells expressed Glut1. In the late proliferating stage, the expression of Glut1 decreased quickly. In the involuting stage, all hemangioma samples didn't express Glut1. All of the samples from the cavernous venous malformations, arteriovenous malformations, capillary malformations and normal skin had no expression of Glut1.

CONCLUSIONSGlut1 may be one of the phenotypes of infantile hemangioma endothelial cells in their development, rather than the inherent character. The expression of Glut1 changes according to the metabolic need of infantile hemangioma cells.

Blood Vessels ; Child ; Child, Preschool ; Endothelium, Vascular ; metabolism ; Glucose Transporter Type 1 ; metabolism ; Hemangioma ; metabolism ; Humans ; Infant ; Phenotype ; Vascular Malformations ; metabolism ; pathology

OBJECTIVEDevelopment and application of a real time fluorescent quantitative PCR (FQ-PCR) assay for detecting WU polyomavirus in children with low respiratory tract infections.

METHODSThe VP2 gene of WU polyomavirus was selected as the detection target, from which the real time primers and probes were designed. The standard curve was established by using recombinant plasmid as template. And the FQ-PCR assay for specific detection of WU polyomavirus was established. The specificity, sensitivity and reproducibility of the method were evaluated. Furthermore, the clinical specimens from children with respiratory tract infections collected in Wenling First People's Hospital were quantitatively detected using this method.

RESULTSIn this study, the FQ-PCR method was established to detect a specific fragment in VP2gene of WU polyomavirus. The standard curve coefficient R2 was 0.998. And this method can detect as low as 50 copies recombinant plasmid. The clinical specimens of sputum and throat swab from children with respiratory tract infections were quantitatively detected using this method. 7 sputum specimens were detected as WU polyomavirus positive in 700 sputum specimens, the positive ratio was 1.00%. No positive specimens were detected in 146 specimens of throat swabs and 846 blood samples from same patient population.

CONCLUSIONThe results indicated that the FQ-PCR assay method established in this study was specific, rapid and sensitive for detecting WU polyomavirus in children with lower respiratory tract infections. The sputum specimen is more suitable to be used for gene detection of WU polyomavirus than throat swab or blood.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Polyomavirus ; isolation & purification ; Real-Time Polymerase Chain Reaction ; methods ; Respiratory Tract Infections ; virology ; Sputum ; virology

OBJECTIVETo investigate the protective effect of tert-butylhydroquinone on bone marrow cells in rats from cytotoxicity induced by benzene in vitro.

METHODSThe bone marrow cells in rats were divided into two groups randomizedly. Cells of the control group were stimulated by 0, 5, 10, 15, 20 mmol/L benzene for 2, 4, 6 hours respectively. Cells of the tBHQ-pretreated group were treated by 100 micromol/L tBHQ for 12 hours followed by the same conditions as the control group. The DNA damage was detected by single cell gel electrophoresis assay (SCGE) and cell apoptosis was examined by flow cytometry. The activities of NAD (P) H: quinone oxidoreductase (NQO1) in bone marrow cells of rats were also measured before benzene treatment in two groups.

RESULTSIn control group, the DNA damage and the apoptosis of bone marrow cells was increased with the growing concentration and time of benzene treatment. The DNA migration and the lengths of DNA migration of the bone marrow cells in the rats under 5, 10, 15, 20 mmol/L benzene treatment in the tBHQ-pretreated group were significantly lower than those in control group at the same time point (P < 0.05). The apoptosis of the bone marrow cells in the rats stimulated by 15, 20 mmol/L benzene for 2 hours and 10, 15, 20 mmol/L benzene for 4 hours as well as 5, 10, 15, 20 mmol/L benzene for 6 hours were also significantly lower than those in control group (P < 0.05). The activities of NQO1 in the bone marrow cells in the rats were increased after tBHQ treatment (P < 0.01) (1.62 +/- 0.16 min(-1).mg(-1) vs. the control group: 0.95 +/- 0.08 min(-1).mg(-1)).

CONCLUSIONThe benzene can induce the DNA damage and the apoptosis of bone marrow cells in rats in a time dependent and dose dependent manner to some extent. The tBHQ can protect the bone marrow cells in rats from the cytotoxicity induced by benzene, which can be partly explained by the increase of the NQO1 activity induced by tBHQ.

Animals ; Apoptosis ; drug effects ; Benzene ; toxicity ; Bone Marrow Cells ; cytology ; drug effects ; enzymology ; Cells, Cultured ; DNA Damage ; drug effects ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; pharmacology ; Hydroquinones ; pharmacology ; Male ; NAD(P)H Dehydrogenase (Quinone) ; metabolism ; Rats ; Rats, Wistar