BACKGROUND:At present, a great quantity of research has shown the effectiveness of traditional Chinese medicine and bone marrow mesenchymal stem cel s for vascular restenosis. However, studies concerning their combined application to restenosis after percutaneous transluminal angioplasty with diabetes mel itus are presently lacking.
OBJECTIVE:To observe the effects of combined application of bone marrow mesenchymal stem cel s and benefiting-Qi nourishing-Yin and dissolving-congestion prescription on restenosis after percutaneous transluminal angioplasty in dogs with diabetes mel itus.
METHODS:A dog model of vascular restenosis with diabetes mel itus was established by bal oon injury of femoral artery and intravenous injection of al oxan. After successful model induction, 22 dog models were randomly divided into three groups:model group (n=6), treatment with Chinese medicine (n=8), and combined treatment with bone marrow mesenchymal stem cel s and Chinese medicine (n=8). Serum vascular endothelial growth factor levels were measured using enzyme-linked immunosorbent assay preoperatively and at 1, 2, 4 and 8 weeks postoperation. Samples of vessels were taken to conduct pathomorphological observation and quantitative analysis of proliferation degree. Tissues, including heart, liver, kidney and pancreatic gland, were col ected to evaluate the safety of stem cel transplantation using hematoxylin-eosin staining at 8 weeks postoperation.
RESULTS AND CONCLUSION:Serum vascular endothelial growth factor levels began to increase at 1 week postoperation in the Chinese medicine group and combined treatment group, at 4 weeks postoperation in the model group compared with preoperation (P<0.05). At al time points, serum vascular endothelial growth factor levels were highest in the combined treatment group, but lowest in the model group (P<0.05). Quantitative analysis of vascular proliferation demonstrated that at 8 weeks postoperation, new intimal area, new intimal/medial areas and stenosis rate were highest in the model group, but lowest in the combined treatment group at 8 weeks postoperation (P<0.05). Safety assessment of stem cel transplantation showed morphological structures of the heart, liver, kidneys and pancreas were normal, no necrosis. In a word, the effects of the combined application of bone marrow mesenchymal stem cel s and benefiting-Qi nourishing-Yin and dissolving-congestion prescription were much pronounced in preventing restenosis after percutaneous transluminal angioplasty in dogs with diabetes mel itus rather than single therapy of Chinese medicine. It is a safe and effective treatment to prevent vascular restenosis after percutaneous transluminal angioplasty in dogs with diabetes mel itus.

The stable and controllable quality of decoction pieces is an important factor to ensure the efficacy of clinical medicine. Considering the dilemma that the existing standardization of processing mode cannot effectively eliminate the variability of quality raw ingredients, and ensure the stability between different batches, we first propose a new strategy for Chinese medicine processing technologies that coupled with individuation processed and cybernetics. In order to explain this thinking, an individual study case about different grades aconite is provided. We hope this strategy could better serve for clinical medicine, and promote the inheritance and innovation of Chinese medicine processing skills and theories.
Chemistry, Pharmaceutical ; methods ; standards ; Cybernetics ; standards ; Drug Therapy ; standards ; Drugs, Chinese Herbal ; chemistry ; standards ; toxicity ; Humans ; Quality Control

Objective To study the expression of immunoglobulin-like transcripts 3 (ⅡT3) and ILT4 in peripheral blood dendritic cells (DC) of kidney transplantation recipients and to analyze its significance in immunity hyporesponsiveness of transplantation. Methods Twenty kidney allograft recipients who were survived more than five years were recruited to two groups: renal function stable groups, chronic rejection groups, and 10 healthy volunteers served as a control group. The peripheral blood mononuclear cells (PBMC) were stimulated with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin 4 (IL-4) and immature DC were obtained. The expression of ILT3 and ILT4 was detected by using flow cytometry. The level of HLA-G5 in serum was determined by using enzyme linked immunosorbent assay. Results ILT3 expression in renal function stable group was increased and decreased in chronic rejection groups as compared with control group (P＜0.05),but ILT4 expression had no significant difference among all groups. HLA-G5 in serum was significantly increased in renal function stable group as compared with other groups. Conclusion Expression of ILT3 and HLA-G was increased in the kidney transplantation recipients with stable renal function and long-term survival, suggesting that they may play an important role in inducing and maintaining periphery immune tolerance.

The specification of decoction pieces and quality uniformity are the important factors to influence the efficacy of clinical medicine. Considering the deficiency of diversity, poor quality uniformity and confusion of decoction pieces specifications, we first propose a new idea of precision decoction pieces (PDP) based on clinical demands and fresh-processed technology. In order to explain the idea, a study case of aconite SUP is provided, including the optimized specification design, processing technology, extraction effects, quality uniformity, and toxic and efficacy variation and so on. The results showed that preparing 5 mm PDP by fresh-cutting is rather simple and practicable, with high efficiency and large yield; then, this technology could significantly decrease the ingredients loss and increase the efficacy components; moreover, it was helpful for achieving the quality uniformity and best extraction effects. This work revealed the quality superiority of PDP, and provided a good strategy and example for the standard of decoction pieces specification and modernization of processing technology.
Aconitum ; chemistry ; Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Particle Size ; Plant Extracts ; chemistry ; pharmacology ; Quality Control

Objective:To compare the two-dimensional electrophoresis(2-DE) maps of rat spinal cord protein extracted by two different solution systems.Methods: Adult rat spinal cord protein was precipitated with 10% trichloracetic acid in acetone and resuspended in 8 mol/L urea plus 4%CHAPS (A solution) or, 5 mol/L urea, 2 mol/L thiourea, 2%CHAPS plus 2%SB3-10 (B solution). One hundred and fifty micrograms of protein was loaded on 18 cm IPG strip holder and run isoelectric focusing electrophoresis as the first dimension, then horizontal SDS-PAGE as the second dimension. Protein spots were visualized by silver stain.Results:There were 1 059 and 1 023 protein spots in each map, of which 790 spots were matched in two maps. There were 269 and 233 spots exclusively extracted by A and B solutions, respectively. Taken together, 1292 different spots were totally obtained by A and B solutions.Conclusion: Integrating protein spots extracted by different solution systems is beneficial for achieving intact 2-DE map of tissues.

Objective To discuss the risk factors attributing intracranial infection following brain injury operation. Methods Difference factors between intracranial infection group and non-infection group following brain injury operations were compared by retrospective analysis. Results Involved in the study were 770 cases in non-infective group and 142 cases (15.6%) in infective group, with the types of infection including nonbacterial meningitis, bacterial meningitis, cerebral ventriclitis and ventrical pus accumulation, cerebral abscess, subdural pus accumulation, subcutaneous or/and subskull flap pus accumulation or osteomyelitis, and incision infections. The rate of bacterial detection was 27.5%.Postoperative infections following brain injury surgery were related to hotter weather, advanced age of patients, severity of traumatic state, multiple operations in short time, successive or double-lateral craniotomies, long-time (＞5 h) operation, microsurgery, operation on skull base or postfossa, cerebral ventricle draining, subcutaneous or epidural hydrops, and emergency operation(P＜0.05). Conclusion It should be given more attention and prophylactic treatments for the brain injury with infective risk factors mentioned above.

Objective:To discuss the topical action characteristics of the biological transmission of moxibustion heat via temperature collection and numerical modeling.Methods:Temperature of moxibustion was measured at multiple points at a distance of 3 cm to obtain the moxibustion temperature field nephograms by the high-accuracy temperature measure array.Finite element analysis was used to imitate the three-dimensional dynamic distribution of temperature in acupoint tissues.Results:Through numerical analysis,the one-dimensional,two-dimensional and three-dimensional distributions of temperature in human acupoint tissues at 5 min of moxibustion were established.The result showed that moxibustion heat mainly transmitted from the surface of the tissue to the internal,and the influence of moxibustion heat decreased with the depth of the tissue.The analysis of the nephograms of acupoint tissue temperature at 5,10,15 and 20 min of moxibustion showed that with the increase of the moxibustion time,the temperature in acupoint tissues constantly rose,and the transmission depth of moxibustion heat also further expanded inside acupoint.Conclusion:By establishing the three-dimensional dynamic model of heat transmission inside acupoint tissues with the biological parameters of human tissues and the temperature values obtained,this study used finite element analysis software ANSYS 14.0 and discovered the rules in the transmission of heat in body tissues during moxibustion,and the features in moxibustion heat transmission (from the proximal to the distant) and heat penetration (from the surface to the internal).This study provides theoretical and experimental support for the application of moxibustion in clinical practice.

To investigate the effects of carbamazepine (CBZ) on the plasma concentrations of valproic acid (VPA) and its toxic metabolite 2-propyl-4-pentenoic acid (4-ene VPA) in epileptic patients, the plasma concentrations of VPA and 4-ene VPA were determined, and the effect of CBZ on pharmacokinetics of VPA was evaluated. All patients had been divided into two groups (VPA group, n = 87; and VPA+CBZ group, n = 19). As compared to VPA group, the combination of CBZ significantly (P < 0.01) decreased the trough concentration of VPA [VPA group, (69.5 +/- 28.8) microg x mL(-1); VPA+CBZ group, (46.3 +/- 25.6) microg x mL(-1)] and does-adjusted VPA trough concentration [VPA group, (4.89 +/- 2.21) microg x mL(-1) x mg(-1) x kg(-1); VPA+CBZ group, (3.14 +/- 1.74) microg x mL(-1) x mg(-1) x kg(-1)]. However, the addition of CBZ did not influence the concentration of 4-ene VPA. The present study revealed that coadministration of CBZ can reduce VPA plasma concentration and may impact VPA clinical effect, therefore therapeutic drug mornitoring of VPA should be used when combined use of CBZ and VPA.
Adolescent ; Adult ; Anticonvulsants ; blood ; pharmacokinetics ; therapeutic use ; Carbamazepine ; blood ; pharmacokinetics ; therapeutic use ; Drug Interactions ; Drug Therapy, Combination ; Epilepsy ; blood ; drug therapy ; Fatty Acids, Monounsaturated ; blood ; Female ; Humans ; Male ; Valproic Acid ; blood ; pharmacokinetics ; therapeutic use ; Young Adult

Objective To evaluate the role of cerebral state index(CSI),burst suppression (BS)and electromyograph(EMG)in monitoring coma/consciousness depth and damage degree of brain. Methods CSM was done in 50 cases with brain injury and coma to analyze its relation with physical reflection,auditory evoked potential(AEP),Glasgow coma score(GCS)and Glasgow outcome scale (GOS).Results As scale range meaning from consciousness to deep coma and to brain death,CSI 0- 100 was positively correlated with coma depth,coma score of GCS and physical reflection.CSI changes under invariable ache stimulation in combination with BS and EMG can accurately estimate prognosis and quantify changes of brain function.Conclusions The quantifiable digit of coma/consciousness depth and damage degree in brain function by CSM can attain real time judgment of dynamic evolvement course of coma and objective guide clinical therapy and assure prognosis,as will change absolutely scoring coma/ consciousness depth and prognosis under current state of artificial diversity and lacking objective evi- dences.

OBJECTIVETo investigate the molecular mechanism of haemophilia B caused by the novel mutation of Arg327Ile (R327I) in FIX gene.

METHODSThe R327I, R327Ala(A), R327Lys(K), R327Asn(N) and a replacement mutant (FIXβFVII), in which FIX β strand 324-329 was replaced by that of FVII 298-303, expression plasmids were constructed with site-directed mutagenesis method based on the wild-type (WT) FIX expression plasmid. The HEK293 cell was transiently transfected, then the activity of FIX (FIX:C) was assayed by one stage method in the conditioned medium, while the FIX:Ag in both the conditioned media and the cell lysates was measured by ELISA. The molecular weight and the semi-quantity of expressed FIX were analyzed by Western blot. Fluorescent protein expression plasmid was constructed to investigate the synthesis and secretion of the FIX R327I mutation in the viable cells.

RESULTSFIX:C of the R327I mutant protein was 4.49% of the level of the WT in the conditioned medium, and the FIX:Ag of the R327I mutant protein in the conditioned medium and the cell lysates was 31.02% and 129.29% compared to that of WT, respectively. The mutation was characterized as cross-reaction material reduced (CRMR). The viable cell fluorescent assays showed that the R327I protein was more in both the viable cells and in lysosome than that of WT. The FIX:C of the R327A, R327K, R327N and FIXβFVII mutants was reduced compared to that of WT, the reduction of FIX:C of FIXβFVII was the most significantly amount among all the mutants in medium. FIX:Ag of all the mutants in the medium, except that the R327K increased, was reduced. The result of Western blot showed that the molecular weight of R327I protein was the same as that of WT, but the amount of the protein was much less compared with WT in the conditioned medium.

CONCLUSIONThe abnormal synthesis and secretion as well as the abnormal function of the R327I mutant protein causes haemophilia B. The residue of R327 as well as the β strand domain of R327 located play important roles of the specific function of FIX.

Factor IX ; genetics ; HEK293 Cells ; Hemophilia B ; genetics ; pathology ; Humans ; Mutagenesis, Site-Directed ; Mutation ; Transfection