Objective To study the effect of autogeneic platelet-rich fibrin (PRF) on proliferation and adipogenic differentiation of human adipose-derived stem cells (ADSCs) in vitro.Methods ADSCs were isolated from adipose tissue obtained from donors undergoing liposuction and were cultured,and underwent identification.ADSCs at passage 3 were divided into three groups:test groups were cultured with 1PRFM and 2PRFM,and control group was cultured without PRF membrane.Then the growth of the cells was observed by inverted microscope.MTT method was used to observe cell proliferation activity at days 1,2,3,4,5,6 and 7 after culture.Adipogenic differentiation of ADSCs was observed and quantified by oil red O staining at days 3,5,7,9,11 and 14.Results Cell proliferation and adipogenic differentiation would be increased with the PRFM,There were significant differences among three groups.Conclusions PRF could significantly promote proliferation and adipogenic differentiation of ADSCs.

The present study is to establish Caco-2/HT29-MTX co-cultured cells and investigate the transport capability of PLGA nanoparticles with different surface chemical properties across Caco-2/HT29-MTX co-cultured cells. PLGA-NPs, mPEG-PLGA-NPs and chitosan coated PLGA-NPs were prepared by nanoprecipitation method using poly(lactic-co-glycolic acid) as carrier material with surface modified by methoxy poly(ethylene glycol) and chitosan. The particle size and zeta potential of nanoparticles were measured by dynamic light scattering. Coumarin 6 was used as a fluorescent marker in the transport of nanoparticles investigated by confocal laser scanning microscopy. The transport of furanodiene (FDE) loaded nanoparticles was quantitively determined by high performance liquid chromatography. Colchicine and nocodazole were used in the transport study to explore the involved endocytosis mechanisms of nanoparticles. Distribution of the tight junction proteins ZO-1 was also analyzed by immunofluorescence staining. The results showed that the nanoparticles dispersed uniformly. The zeta potential of PLGA-NPs was negative, the mPEG-PLGA-NPs was close to neutral and the CS-PLGA-NPs was positive. The entrapment efficiency of FDE in all nanoparticles was higher than 75%. The transport capability of mPEG-PLGA-NPs across Caco-2/HT29-MTX co-cultured cells was higher than that of PLGA-NPs and CS-PLGA-NPs. Colchicine and nocodazole could significantly decrease the transport amount of nanoparticles. mPEG-PLGA-NPs could obviously reduce the distribution of ZO-1 protein than PLGA-NPs and CS-PLGA-NPs. The transport mechanism of PLGA-NPs and mPEG-PLGA-NPs were indicated to be a combination of endocytosis and paracellular way, while CS-PLGA-NPs mainly relied on the endocytosis way. PEG coating could shield the surface charge and enhance the hydrophilicity of PLGA nanoparticles, which leads mPEG-PLGA-NPs to possess higher anti-adhesion activity. As a result, mPEG-PLGA-NPs could penetrate the mucus layer rapidly and transport across Caco-2/HT29-MTX co-cultured cells.
Biological Transport ; Caco-2 Cells ; Chitosan ; chemistry ; Coated Materials, Biocompatible ; chemistry ; Coculture Techniques ; Drug Carriers ; Furans ; administration & dosage ; chemistry ; metabolism ; HT29 Cells ; Heterocyclic Compounds, 2-Ring ; administration & dosage ; chemistry ; metabolism ; Humans ; Lactic Acid ; chemistry ; Nanoparticles ; Particle Size ; Polyethylene Glycols ; chemistry ; Polyglycolic Acid ; chemistry ; Zonula Occludens-1 Protein ; metabolism

Objective To explore the dynamic changes of the cellular immune function in severe infection after liver transplantation, and to guide the individualized immunology adjustment. Methods 378 cases of livertransplantation were analyzed retrospectively. Seventy-four cases (infection group) suffered serious infection, including 54 cases cured (cure group), 20 cases died (death group). Fifty cases without infection and rejection were randomly selected as control group (stable group). According to the individualized adjusting proposal of immunosuppressants, 74 patients with severe infection were divided into two groups: traditional (T) group and individualized (Ⅰ) group. The general condition, recovery rate and change of cellular immune function pre- and post-treatment were analyzed. Results The preoperative MELD score and the intraoperative blood loss in infection group were significantly higher than stable group, and those in death group were higher than in cure group. CD4+ T lymphocyte counts and lymphocyte counts in stable group were increased significantly from first week post-operation to discharge. The two indicators in infection group at first week postoperation and the onset of infection were lower than in stable group (P<0. 01). In cure group after infection was controlled the two indicators were higher than at first week post-operation and the onset of infection (P<0. 01), while in death group they were reduced up to death (P<0. 05). There was no significant difference in age, preoperative MELD score and the immune function indicators both at first week post-operation and the onset of infection between T group and Ⅰ group, except the intraoperative blood loss in Ⅰ group was greater than in T group. The recovery rate in Ⅰ group (90. 5 %)was higher than in T group (66.0 %). Conclusion Individualized adjustments of immunosuppressants guided according to the dynamic changes of cellular immune function helped to improve the prognosis of severe infection after liver transplantation.

OBJECTIVETo analyze the epidemiologic characteristics of TCM syndrome distribution and season correlation in hypertension patients.

METHODSClinical materials regarding population, symptoms and signs, etc. of 698 hypertension patients were collected, and statistical analysis on them was conducted.

RESULTSTCM syndrome type revealed in patients, in order of frequency from high to low, were: Gan-fire flaming-up syndrome, yin-deficiency and yang-excess syndrome, phlegm-dampness accumulation syndrome, both yin-yang deficiency syndrome, and others. The incidence of various syndromes was higher in the winter/spring and rather low in the summer, and Gan-fire flaming-up syndrome was positively related to the spring and negative to the summer.

CONCLUSIONSThe syndrome types of hypertension patients are chiefly the Gan-fire flaming-up syndrome, yin-deficiency and yang-excess syndrome, phlegm-dampness accumulation syndrome, and both yin-yang deficiency syndrome; Gan-fire flaming-up syndrome is the dominance; their incidence is higher in the winter/spring and lower in the summer.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Female ; Health Surveys ; Humans ; Hypertension ; diagnosis ; epidemiology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Seasons ; Young Adult

OBJECTIVE: To observe the expression pattern of caspase-3 and HCLS1-associated protein X-1 (HAX-1) at different time after cerebral contusion in rat, and explore the new method for estimating the injury interval. METHODS: The cerebral contusion model was established using adult SD male rats. Then the rats were randomly allocated into 8 groups: 2 h, 6 h, 12 h, 1 d, 3 d, and 7 d after cerebral contusion, sham-operation and normal control. Expression of caspase-3 and HAX-1 protein after cerebral contusion in rat was detected by Western blotting. Laser scanning confocal microscope was used to observe the number of HAX-1 positive cells and TUNEL-stained cells after cerebral contusion. RESULTS: The expression of caspase-3 increased parallelly with the time after cerebral contusion and reached the peak value on 3 d. The expression of caspase-3 decreased gradually and still maintained a high level expression on 7 d (P < 0.05). The expression of HAX-1 positive cell went up after injury, and reached the peak value at 6 h (P < 0.05), then turned down gradually after 12 h and went out of detection after 3 d. The number of TUNEL-stained cells increased obviously at 2 h and reached the peak value on 3 d. The number of TUNEL-stained apoptotic cells decreased gradually and still maintained a high level expression on 7 d (P < 0.05). CONCLUSION The expression of caspase-3 and HAX-1 after cerebral contusion has time sequential regularity, which may provide new evidence for forensic diagnosis of cerebral contusion interval.
Animals ; Blotting, Western ; Brain Injuries/pathology* ; Carrier Proteins/metabolism* ; Caspase 3/metabolism* ; Cerebellum/pathology* ; In Situ Nick-End Labeling ; Intracellular Signaling Peptides and Proteins ; Male ; Rats ; Rats, Sprague-Dawley

The authors designed to separate, purify and determine the monosaccharide composition of the polysaccharide from Cordyceps militaris, and study its effect on reverse cholesterol transport in vivo by isotope tracing assay. Polysaccharides were separate and purify by ion exchange column Q-sepharose Fast Flow and size exclusion column Sephacryl S200HR; the molecular weight and monosaccharide composition of the polysaccharides were determined by high performance gel permeation chromatography and high performance liquid chromatography coming with pre-column derivation, respectively. Finally, three purified polysaccharides CMBW1, CMBW2 and CMYW1 were obtained, their total carbohydrate contents were 87%, 89%, 95%, respectively; their protein contents were 6.5%, 1.3%, 2.8%, respectively; their molecular weights were 772.1, 20.9, 13.2 kDa, respectively; CMBW1 was composed of mannose, glucosamine, rhamnose, glucuronic acid, glucose, galactose and arabinose with a molar ratio of 7.25: 0.17: 1.29: 0.23: 6.30: 11.08: 0.79; CMBW2 was composed of mannose, glucosamine, galactose and arabinose with a molar ratio of 2.40: 0.16: 2.92: 0.24; CMYW1 was composed of mannose, glucosamine, glucuronic acid and glucose with a molar ratio of 0.59: 0.57: 0.45: 25.61. Polysaccharide at 50 mg x kg(-1) could significantly improve the transport of 3H- cholesterol to blood and excretion from feces. All of the three purified polysaccharides CMBW1, CMBW2 and CMYW1 were heteropolysaccharide; and they could improve reverse cholesterol transport in vivo, the underlying mechanisms are being studied.
Animals ; Biological Transport ; drug effects ; Cholesterol ; metabolism ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Cordyceps ; chemistry ; Mice ; Monosaccharides ; analysis ; isolation & purification ; Polysaccharides ; chemistry ; isolation & purification ; pharmacology ; Tritium

The aim of this study is to establish an HPLC method for simultaneous determinations of mifepristone and its metabolites, mono-demethylated mifepristone, di-demethylated mifepristone and C-hydroxylated mifepristone in plasma and to evaluate the pharmacokinetic characteristics of mifepristone tablet. Twenty healthy female Chinese subjects were recruited and a series of blood samples were collected before and after 0.25, 0.5, 1.0, 1.5, 2.0, 4.0, 8.0, 12.0, 24.0, 48.0, 72.0 and 96.0 hours administration by a single oral dose of 75 mg mifepristone tablet. Mifepristone and its three metabolites were extracted from plasma using ethyl acetate and determined by high performance liquid chromatography. The main pharmacokinetic parameters of mifepristone and its metabolites, including Cmax, tmax, MRT, t(1/2), V, CL, AUC(0-96 h) and AUC(0-infinity), were calculated by Drug and Statistical Software Version 2.0. The simple, accurate and stable method allows the sensitive determinations ofmifepristone and its metabolites in human plasma up to 4 days after oral administration of 75 mg mifepristone tablet and the clinical applications of their pharmacokinetic studies.

OBJECTIVETo explore the tumor growth inhibition of gamma secretase inhibitor MRK003 on human multiple myeloma xenograft mice by inhibition of AKT and Notch1 expression.

METHODSNOD/SCID mice were injected with human multiple myeloma cell lines RPMI8226 to establish a xenograft mouse model. Mice were randomized into two groups：the experimental group were injected with MRK003 at a dose of 5 mg× kg⁻¹×d⁻¹ for 14 days; the inhibitor was replaced by an equal saline in the control group. Mice were sacrificed by cervical dislocation on the next day after the last injection and tumor tissue was removed to detect the expression of Notch1 and AKT by immunohistochemistry.

RESULTSAfter subcutaneous injection with RPMI8226, mice had tumor formation in 5-7 days and the largest tumor block in 10-12 days. Before RPMI8226 injection, the mean sizes of tumor block in the experimental and the control groups were 509.2 mm³, 511.2 mm³(P>0.05). 9 days after injection, the mean sizes of tumor tissue in the experimental and the control groups were 636.6 mm³, 691.2 mm³(P<0.01). On the next day after the last injection, the tumor sizes of the experimental and the control groups were 683.5 mm³ and 1798.7 mm³(P<0.01). The size of tumor block in the experimental group was significantly smaller than that of the control group(P<0.01). Immunohistochemistry staining showed that the positive expression rates of Notch1(11.1%, P<0.01) and AKT(13.3%, P<0.01) in experimental group were significantly decreased compared with the control group(Notch1: 95.6%; AKT: 93.3%). Western blot results showed that Notch1 and AKT protein in experimental group were significantly lower than those in the control group.

CONCLUSIONMRK003 could inhibit the tumor growth of human multiple myeloma xenograft mice by downregulated expression of Notch1 signaling pathway.

Amyloid Precursor Protein Secretases ; antagonists & inhibitors ; Animals ; Cell Line, Tumor ; Cyclic S-Oxides ; pharmacology ; Humans ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Multiple Myeloma ; drug therapy ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Receptor, Notch1 ; metabolism ; Signal Transduction ; drug effects ; Thiadiazoles ; pharmacology ; Xenograft Model Antitumor Assays

OBJECTlVE To study the Iong term toxicity of vinoreIbine tartrate(NVB)on rat immune and hematopoietic systems pathoIogicaIIy. METHODS SD Rats were randomIy divided into 4 groups:normaI controI group and NVB 5.0,10.0,and 20.0 mg·m-2 groups,each group containing 6 maIe and femaIe rats. The rats in NBV groups were administered different concentrations of NVB by intravenous drip on the 1st and 8th days,21 da cycIe,for 4 cycIes. On the 14th day after the Iast administration, white bIood ceIIs(WBC),neutrophiI(Neut),Iymphocytes(Lym),red bIood ceIIs(RBC)and reticuIo-cyte‰(RET‰)were detected by ADVIA2120 hematoIogy anaIyzer. Thymus,sternum marrow,spIeen and mesenteric Iymph nodes were observed by histopathoIogicaI examination. The thymus and spIeen were preciseIy weighed to obtain the reIative organ coefficients. Bone marrow smears were made for counting and cIassification. RESULTS Compared with normaI controI group,WBC,Neut,Lym,RBC and RET% of peripheraI bIood of NVB 5,10 and 20 mg·m-2 groups were decreased(P﹤0.05,P﹤0.01). The Neut vaIue of maIe rats was(2.35±0.56)×109·L-1 in normaI controI group,but was reduced to (1.66±0.44),(0.67±0.22)and(0.20±0.02)×109·L-1(P﹤0.05,P﹤0.01)in NVB 5,10 and 20 mg·m-2 groups. The Neut vaIue of femaIe rats was(1.26± 0.27)× 109 L-1 in normaI controI group,but was reduced to(1.14±0.56),(0.47±0.13)and(0.21±0.08)×109 L-1(P﹤0.05,P﹤0.01)in NVB 5,10 and 20 mg·m-2 groups. The resuIts of counting and cIassification of bone marrow smears showed that the myeIoid ceII ratio decreased(P﹤0.05,P﹤0.01). The myeIoid ceII ratio of maIe rats was(42.7±6.1)% in normaI controI group,but was reduced to(28.8±5.3)%,(22.0±3.2)% and(18.9±3.9)% in NVB 5,10 and 20 mg·m-2 groups. The myeIoid ceII ratio of femaIe rats in normaI controI group was(35.4±3.0)%, but was reduced to(31.2±4.7)%,(22.9±6.7)% and(20.8±4.2)% in NVB 5,10 and 20 mg·m-2 groups. The thymus coefficient was reduced(P﹤0.05,P﹤0.01). The thymus coefficient of maIe rats in normaI controI group was 0.36±0.04,but was reduced to 0.31±0.06,0.18±0.03 and 0.08±0.01 in NVB 5,10 and 20 mg·m-2 groups. The thymus coefficient of femaIe rats in normaI controI group was 0.29±0.06,but was reduced to 0.25±0.06,0.19±0.06 and 0.07±0.01 in NVB 5,10 and 20 mg·m-2 groups. Histopatho-IogicaI examination showed that thymus was atrophiedand bone marrow was suppressed. SpIeen com-pensatory extrameduIIary hematopoietic ceIIs were increased in NVB 5.0,10.0 and 20.0 mg·m-2 groups (maIe and femaIe)to different degrees,but the mesenteric Iymph nodes of NVB groups showed no sig-nificant pathoIogicaI changes. CONCLUSlON NVB has immune and hematopoietic toxicity on SD rats, as is showed by thymic atrophy and bone marrow suppression.

Objective To develop a supporting pole for medical detachable camouflage net at field conditions to enhance the timeliness of battlefield camouflage of deployable units such as medical tent.Methods The pole had a dumbbell-shaped,hollow and columnar structure,which was composed of a base,a pole body and a terminal disc.The pole body consisted of internal and external parts.The external part had a vertical opening at its top and screw thread at its side wall,which was equipped with a binding bolt.The terminal disc had a circular structure and a 20 cm outer diameter,which was fixed 10 cm under the top of the internal pole to support the net.The base had a center hole and a sleeve to hold the lower part of the external pole to immobilize the supporting pole.Results The supporting pole decreased the deployment time of the camouflage net from 5 min to 2 min and the withdrawal time from 4 min to 1.5 min,and enhanced the timeliness of all-element deployment of mobile medical unit during field practical training.Conclusion The supporting pole meets the tactical requirements for deployment,storage,transport and robustness,and thus is worthy promoting in medical unit.