Objective To explore the mechanism of Angelica Sinensis polysaccharide(ASP)promoting donor bone marrow transplantation(BMT)to reconstruct hematopoietic function of receptor mice by regulating bone marrow stromalcells(BMSCs).Methods Bone marrow mononuclear cells(BMMNCs)of male C57BL/6J mice aged 8-10 weeks were separated,purified and transplanted into female receptor mice of the same age.On the ninth day,receptor mice BMMNCs were separated,purified and transplanted again into female receptor mice.The transplanted receptor mice were divided into control group:sham irradiation;Irradiation(IR)group:a whole-body irradiation with a total dose of 8.0 Gy X-ray;BMT group:the receptor mice treated in the same way as the IR group and transplanted BMMNCs(5×106 cells)from male donor via the tail vein;BMT+ASP group:the receptor mice treated in the same way as the BMT group,and injected ASP[100 mg/(kg·d)×9]by intraperitoneal route from the first day of transplantation.Changes in body weight and survival rate of mice were recorded during modeling,receptor mice BMMNCs were collected to detect sex-determining region of Y(SRY)gene after building model,peripheral blood indexes,the number of BMMNCs in femur and histopathology of bone marrow were detected;BMSCs in receptor mice was separated and purified,BMSCs adhesion ability was observed,proliferation ability was detected by 5-ethynyl-2-deoxyuridine(EdU);The level of reactive oxygen species(ROS),the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in BMSCs were detected;The levels of granulocyte-macrophage colony-stimulating factor(GM-CSF),stem cell factor(SCF),insulin-like growth factor 1(IGF-1)in culture supernatant of BMSCs were determined,CFU-Mix was counted after BMMNCs co-cultured with receptor BMSCs in each group for 48 hours;The expression of Notch signaling pathway related genes(Notch1,Jagged1,Hes1)in BMMNCs were measured by Real-time PCR.Results All mice in IR group were died,the body weight loss in BMT+ASP group was not obvious.The SRY gene was detected in the receptor female mice BMMNCs.Peripheral blood indexes and the number of BMMNCs were not significantly decreased in BMT+ASP group receptor mice,and bone marrow histopathological injury was reduced.ASP promoted the proliferation of BMSCs,decreased the contents of ROS and MDA,and increased the activity of SOD in BMSCs.ASP promoted the secretion of SCF,GM-CSF and IGF-1 in BMSCs,and increased CFU-Mix yield of BMMNCs co-cultured with receptor BMSCs.ASP increased the expression of Notch1,Jagged1 and Hes1 mRNA in BMMNCs.Conclusion The mechanism of ASP promoting receptor hematopoietic function reconstruction is related to reducing the oxidative stress damage of hematopoietic microenvironment,improving the secretion of hematopoietic growth factors in BMSCs,and regulating Notch signaling pathway.

Objective:To observe the effect of different doses of Fufang Huangbaiye Tuji asin the treatment onof the inflammatory response in healing process for of skin with deep Ⅱ degree burn. Methods in healing process. Methods:The 120 cses patients with deep Ⅱ degree burn of fire-toxin injuring fluid syndrome diagnosed in the affiliated hospital of Chengde Medical University between June 2019 and March 2020 were randomly divided into control group，low -dose treatment group and high -dose treatment group，with 40 cases in each group and once. They got a dressing change perevery day. Control group was locally administered with lodophor solution 35 mL per 1% on the body surface area. Low-dose treatment group was locally administered with compound cortex phellodendri fluid 17.5 mL per 1% on the body surface area，while high-dose treatment group was locally administered with compound cortex phellodendri fluid 35 mL per 1% on the body surface area. Observe theThe inflammatory reaction of wound surface in each group onwas observed at admission and after treatment. The pathological changes of each groupsgroup were observed， and determination of nuclear factor kappa-B（NF-κB） p65 expression inon the wound surface was determined by immunohistochemistry on the 4th day after the treatment. The levels of interleukin（IL）-2，IL-8 and tumor necrosis factor（TNF）-α in wound tissue were measured with ELISA and Bacterial culture and count were performed in each group on the 4^th，10^th and 21^st daydays after treatment. The levels of IL-2，IL-8 and TNF-α in wound tissue were measured with ELISA. Results:There was no significant difference in the degree of wound inflammation in each group at admission，and the degree of relief after treatment was positively correlated with the treatment time. At the simultaneous phase point，the inflammatory reaction was severest in control group，which was followed by low-dose treatment group and high-dose treatment group. Bacterial growth were observed on the 4^th day in control group，which was found in low-dose and high-dose treatment groups on the 10^th day，the detection rates of Staphylococcus aureus and Pseudomonas aeruginosa were the highest. Compared with control group，the mean integrated optical density of NF-κB p65 in wound tissue decreased markedly in low-dose and high-dose treatment groups on the 4th day after treatment（P<0.05），the bacterial count decreased significantly in low-dose and high-dose treatment groups on the 10^th and 21^st days after treatment（P<0.05），and the levels of IL-2，IL-8 and TNF-α in wound tissue decreased markedly in low-dose and high-dose treatment groups on the 4^th，10^th and 21^st days after treatment（P<0.05），with statistically significant differences between low-dose and high-dose treatment groups（P<0.05）. Histopathological examination showed that inflammatory granulocytes and edema were improved in low-dose and high-dose treatment groups compared with control group，with a more significant performance in high-dose treatment group. Conclusion:The external application of compound cortex phellodendri fluid can reduce thebacterial growth of bacteria in on the wound surface，which may reduce the inflammatory reaction by inhibiting the production and release of inflammatory mediators，with a certain dose-effcteffect relationship，and is worth clinical promotion.

Objective:To study the intervention effect and underlying mechanism of Fufang Huangbaiye Tuji （FFHBY） on skin with deep Ⅱ degree burn wound. Method:Patients with deep Ⅱ degree burn of fire-toxin injuring fluid syndrome diagnosed in the Affiliated Hospital of Chengde Medical University from June 2019 to June 2020 were randomly divided into a control group （iodophor solution， 35 mL per 1% body surface area）， a low-dose treatment group （FFHBY， 17.5 mL per 1% body surface area）， and a high-dose treatment group （FFHBY， 35 mL per 1% body surface area）， 40 cases in each group. The patients in each group were treated correspondingly with dressing chance once per day. The pathological changes of the wound were observed on the 14th day after treatment. Wound symptoms and signs in each group before treatment and on the 7th， 14th， and 21st days after treatment were quantified， and the clinical efficacy on the 21st day after treatment was evaluated. Wound healing rates in each group were calculated on the 7th， 14th， and 21st days after treatment. The levels of vascular endothelial growth factor （VEGF）， fibroblast growth factor （FGF）-2， FGF-7， epidermal growth factor （EGF）， interleukin （IL）-10， tumor necrosis factor （TNF）-α， and Caspase-3 in wound tissues were measured with enzyme-linked immunosorbent assay （ELISA）. Nuclear factor kappa-B （NF-κB） p65 expression in wound surface was detected by immunohistochemistry. The apoptosis rate in wound tissues was determined by the TdT-mediated dUTP-biotin nick end labeding assay （TUNEL） method. Result:There was no significant difference in scores of symptoms and signs among groups before treatment. Compared with the control group， the treatment groups showed no significant difference in wound healing rates on the 7th day after treatment and increased healing rates on the 14th and 21st day after treatment（P<0.05）. The clinical efficacy in the treatment groups was superior to that in the control group on the 21st day after treatment. Additionally， the treatment groups also showed decreased scores of local symptoms and signs， increased levels of VEGF， FGF-2， FGF-7， EGF， and IL-10， and dwindled apoptosis rate and levels of Caspase-3， TNF-α， and NF-κB p65 expression in wound tissues on the 7th，14th and 21st day after treatment （P<0.05）. The high-dose treatment group was superior to the low-dose treatment group in the above indicators （P<0.05）. Histopathological examination showed that inflammatory cell infiltration was relieved in the treatment groups as compared with that in the control group， and the high-dose treatment group exhibited superior efficacy. Conclusion:FFHBY had an obvious therapeutic effect on deep Ⅱ degree burn. It could promote wound healing by up-regulating the level of growth factors， improving inflammatory response， and inhibiting cell apoptosis in a dose-dependent manner.

BACKGROUND: Umbilical cord mesenchymal stem cells (UC-MSCs) are a group of cells that have self-renewal, highly proliferative and multidrug differentiation potential. The properties of UC-MSCs and their tumor tropism make them an ideal tool for glioma cell therapy. These cells can act by paracrine or as a delivery system for genes and drugs. It has been demonstrated that UC-MSCs can inhibit the growth of glioma and improve the survival after transplantation into the brain. OBJECTIVE: To summarize the molecular mechanisms and safety of UC-MSCs in the treatment of glioma and to provide a useful reference for further research. METHODS: We searched the PubMed and CNKI databases from 2000 to 2017 with the English terms of "glioma; umbilical cord mesenchymal stem cells" and the Chinese terms of "glioma; umbilical cord mesenchymal stem cells; safety; molecular mechanism". Based on the inclusion and exclusion criteria, 55 articles were finally reserved for review. RESULTS AND CONCLUSION: UC-MSCs have obvious effect on treating glioma. These cells can treat glioma through homing mechanism and paracrine mechanism as gene carrier and co-culture. Moreover, UC-MSCs have certain safety in the treatment of glioma.

Objective To analyse the genetic variability of EG95 sequences and provide guidance for EG95 vaccine application against Echinococcus granulosus (E. granulosus). Methods We analysed EG95 polymorphism by collecting total 97 different E. granulosus isolates from 12 different host species that originated from 10 different countries. Multiple sequence alignments and the homology were performed by Lasergene 1 (DNASTAR Inc., Madison, WI), and the phylogenetic analysis was performed by using MEGA5.1 (CEMI, Tempe, AZ, USA). In addition, linear and conformational epitopes were analysed, including secondary structure, NXT/S glycosylation, fibronectin type III (FnIII) domain and glycosylphosphatidylinositol anchor signal (GPI-anchor). The secondary structure was predicted by PSIPRED method. Results Our results indicated that most isolates overall shared 72.6–100% identity in EG95 gene sequence with the published standard EG95 sequence, X90928. However, EG95 gene indeed has polymorphism in different isolates. Phylogenetic analysis showed that different isolates could be divided into three subgroups. Subgroup 1 contained 87 isolates while Subgroup 2 and Subgroup 3 consisted of 3 and 7 isolates, respectively. Four sequences cloned from oncosphere shared a high identity with the parental sequence of the current vaccine, X90928, and they belonged to Subgroup 1. However, in comparison to X90928, several amino acid mutations occurred in most isolates besides oncosphere, which potentially altered the immunodominant linear epitopes, glycosylation sites and secondary structures in EG95 genes. All these variations might change their previous antigenicity and thereby affecting the efficacy of current EG95 vaccine. Conclusions This study reveals the genetic variability of EG95 sequences in different E. granulosus isolates, and proposed that more vaccination trials would be needed to test the effectiveness of current EG95 vaccine against distinct isolates in different countries.

The present study aimed at isolation and purification of the bioactive terpenoids from the herb of Leonurus japonicus by chromatographic separations such as silica gel, sephadex LH-20 and C18 reversed phase silica gel, as well as preparative HPLC. As a result, leojaponic acids A (1, C17H24O4) and B (2, C18H26O4), two homologous terpenoids, together with (-)-loliolide (3), 1-(3-ethylphenyl) ethane-1, 2-diol (4) and dibutyl phthalate (5), were isolated from the EtOH extract of L. japonicus. All the chemical structures of the isolates were elucidated on the basis of 1D and 2D NMR analyses. Compounds 1 and 2 were new terpenoids, and Compounds 3 and 4 were isolated and identified for the first time from this plant. In addition, the α-glucosidase and tyrosinase inhibitory activity of the new compounds were evaluated.
Enzyme Inhibitors ; chemistry ; isolation & purification ; Fruit ; chemistry ; Glucosidases ; analysis ; antagonists & inhibitors ; Leonurus ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Extracts ; chemistry ; isolation & purification ; Terpenes ; chemistry ; isolation & purification

Toxicity of different processed was evaluated Polygoni Multiflori Radix by determining the hepatotoxic potency for selecting processing technology. Process Polygoni Multiflori Radix using high pressure steamed, Black Bean high pressure steamed, atmospheric steamed for different time. Using normal human hepatocytes (L02) as evaluation model, hepatotoxic potency as index to evaluate hepatotoxic potency of different processed Polygoni Multiflori Radix. Analysis chemical composition of some processed products by UPLC-MS. Hepatotoxic bioassay method cloud evaluate the toxicity of different Polygoni Multiflori Radix samples. Different processing methods can reduce the toxicity of Polygoni Multiflori Radix, high pressure steamed three hours attenuated was better. Different processing methods have different effects on chemical constituents of Polygoni Multiflori Radix. Comparing with crude sample, the contents of gallic acid, 2,3,5,4-tetrahydroxyl diphenylethylene-2-O-glucoside, emodin-8-O-beta glucoside and emodin were decreased in processed products with 3 kinds of different methods. The change trend of 2,3,5,4-tetrahydroxyl diphenylethylene-2-O-glucoside content was similar with hepatotoxic potency. Different processing methods can reduce the toxicity of Polygoni Multiflori Radix. Processing methods and time attenuated obvious impact on toxicity. Recommended further research on the attehuated standard control of Polygoni Multiflori Radix concocted.
Biological Assay ; Cell Line ; Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Fallopia multiflora ; chemistry ; toxicity ; Hepatocytes ; drug effects ; Humans ; Plant Roots ; chemistry ; toxicity

BACKGROUNDCell transplantation has great potential for promoting endothelial repair and reducing the complications of percutaneous coronary intervention (PCI). The aim of this study was to investigate the effect of transplantation of human umbilical cord blood endothelial progenitor cells (EPCs) on injured arteries.

METHODSUmbilical cord blood mononuclear cells were obtained from post-partum lying-in women, and EPCs were isolated, cultured, expanded and identified by immunofluorescence. The carotid arterial endothelium of New Zealand white rabbits was injured by dilatation with a 3F balloon, and the EPCs were injected into the lumen of the injured artery in the transplanted group (n = 16), while an equal volume of phosphated buffered saline (PBS) was injected into the control group after balloon injury (n = 16). The animals were sacrificed after either 2 or 4 weeks, and the grafted cells were identified by double immunofluorescence staining with human nuclear antigen (HNA) and CD31 antibodies. Arterial cross sections were analyzed by pathology, immunohistochemistry and morphometry to evaluate the reparative effects of EPCs. Proliferating cell nuclear antigen (PCNA) and transforming growth factor (TGF)-β1 mRNA expression were detected by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSFluorescence-labeled EPCs were found in the neointima. The neointimal area and the neointimal/medial area ratio were significantly lower in the transplanted group than in the control group (P < 0.05). von Willebrand factor (vWF) immunohistostaining showed more VWF-positive cells in the transplanted animals than in the controls (8.75 ± 2.92 vs. 4.50 ± 1.77, P < 0.05). Compared with the control group, the transplanted group had lower expression of PCNA mRNA (0.67 ± 0.11 vs. 1.25 ± 0.40, P < 0.01) and higher expression of TGF-β1 mRNA (1.10 ± 0.21 vs. 0.82 ± 0.07, P < 0.05).

CONCLUSIONSEPCs derived from human umbilical cord blood were successfully transplanted into injured vessels. The transplanted EPCs inhibited neointimal hyperplasia and promoted vascular re-endothelialization.

Animals ; Carotid Artery Injuries ; immunology ; pathology ; therapy ; Cell Differentiation ; Cells, Cultured ; Cytokines ; genetics ; Endothelial Cells ; cytology ; physiology ; Fetal Blood ; cytology ; Humans ; Hyperplasia ; Male ; Neointima ; pathology ; Proliferating Cell Nuclear Antigen ; genetics ; RNA, Messenger ; analysis ; Rabbits ; Stem Cell Transplantation ; Transforming Growth Factor beta1 ; genetics

Reasonable sampling scheme is the important basis for establishing reliable population pharmacokinetic model. It is an effective method for estimation of population pharmacokinetic parameters with sparse data to perform population pharmacokinetic analysis using the nonlinear mixed-effects models. We designed the sampling scheme for amlodipine based on D-optimal sampling strategy and Bayesian estimation method. First, optimized sample scenarios were designed using WinPOPT software according to the aim, dosage regimen and visit schedule of the clinical study protocol, and the amlodipine population model reported by Rohatagi et al. Second, we created a NONMEM-formatted dataset (n = 400) for each sample scenario via Monte Carlo simulation. Third, the estimation of amlodipine pharmacokinetic parameters (clearance (CL/F), volume (V/F) and Ka) was based on the simulation results. All modeling and simulation exercises were conducted with NONMEM version 7.2. Finally, the accuracy and precision of the estimated parameters were evaluated using the mean prediction error (MPE) and the mean absolute error (MAPE), respectively. Among the 6 schemes, schemes 6 and 3 have good accuracy and precision. MPE is 0.1% for scheme 6 and -0.6% for scheme 3, respectively. MAPE is 0.7% for both schemes. There is no significant difference in MPE and MAPE of volume among them. Therefore, we select scheme 3 as the final sample scenario because it has good accuracy and precision and less sample points. This research aims to provide scientific and effective sampling scheme for population pharmacokinetic (PK) study of amlodipine in patients with renal impairment and hypertension, provide a scientific method for an optimum design in clinical population PK/PD (pharmacodynamics) research.
Adult ; Age Factors ; Alanine Transaminase ; blood ; Amlodipine ; pharmacokinetics ; pharmacology ; Antihypertensive Agents ; pharmacokinetics ; pharmacology ; Bayes Theorem ; Body Weight ; Calcium Channel Blockers ; pharmacokinetics ; pharmacology ; Humans ; Hypertension ; metabolism ; Metabolic Clearance Rate ; Middle Aged ; Models, Biological ; Monte Carlo Method ; Nonlinear Dynamics ; Renal Insufficiency ; metabolism ; Software

OBJECTIVETo study the clinical value of magnetic resonance spectroscopy (MRS) image in stereotactic biopsy for brain lesion.

METHODSFrom April 2008 to April 2010, 126 cases (72 male and 54 female, aged from 10 to 82 years, mean 45 years) of brain lesion which were difficult to diagnose were divided into two groups by random number table, 62 cases were executed for MRI-guided frameless stereotactic biopsy (MRI group), 64 cases were executed for MRI and MRS-guided frameless stereotactic biopsy (MRS group). Operation used MRI and Three-dimensional MRS image to locate, and used frameless CAS-R-2 robots to carry out the positioning operating.

RESULTSNo surgery-related deaths and infections. Pathological diagnosis was 106 cases of brain tumors, 6 cases of inflammatory disease, 4 cases of tumor-like demyelinating disease and multiple sclerosis, 3 cases of neurodegenerative disease, 7 cases failed to obtain positive pathological diagnosis. The total rate of positive diagnosis was 94.4%, the positive rate in MRS-guided stereotactic biopsy group was 98.4% (63/64), the positive rate of conventional MRI-guided biopsy group was 90.3% (56/62), and there was statistically significant difference between the two groups (χ(2) = 3.92, P = 0.047). Four cases presented with postoperative complications, the complication rate was 3.2% (4/126); the complications were cerebral hemorrhage associated with aphasia, epilepsy, subcutaneous hematoma, gastrointestinal bleeding, which were improved after treatment.

CONCLUSIONSMRS-guided stereotactic biopsy group has a higher positive rate than MRI-guided stereotactic biopsy group, indicating that this method can improve the positive rate of diagnosis, and thus will help to formulate treatment plan for brain lesion.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biopsy ; methods ; Brain ; pathology ; Brain Diseases ; pathology ; Brain Neoplasms ; pathology ; Child ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Young Adult