OBJECTIVE: To analyze and discuss four methods of calculating likelihood ratio of DNA mixture. METHODS: In the case with CNAS-T0757 proficiency testing in 2013, the likelihood ratios were calculated and compared among four methods, including unrestricted combinatorial method, Clayton's method, p2 principle method, and recommendations from ISFG. RESULTS: The likelihood ratios were maximum by Clayton's method and recommendations from ISFO, followed by result of the unrestricted combinational method. The minimum likelihood ratio was obtained by p2 principle. CONCLUSION The unrestricted combinational method could give fUrthest consideration to both information preservation and appraiser protection.
DNA/genetics* ; DNA Fingerprinting ; Humans ; Likelihood Functions

The objective of the present study was to establish a method based on principal component analysis (PCA) for the study of transdermal delivery of Chinese medicinal formulae, and to choose the best penetration enhancers for Qingfei Xiaocuo gel depend on this method. Using improved Franz type diffusion cell and excised rat skin in vitro as transdermal barrier, the receptive solution fingerprint was established by HPLC, harvesting the areas of the common peaks in the fingerprint, then the total factor scores of the concentrations at different times were calculated using PCA and were employed instead of the concentrations to compute the cumulative amounts (Q12) and enhancement ratio (ER), the latter of which were considered as the indexes for optimizing penetration enhancers. Compare to the control group, the ER of the other groups increased significantly and furthermore, 2.5% azone with 2.5% menthol manifested the best effect. PCA represent most information in the receptive solution, the method above could choose the best penetration enhancers, it could be a reference for the study of transdermal delivery of Chinese medicinal formulae.
Administration, Cutaneous ; Animals ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Gels ; In Vitro Techniques ; Male ; Medicine, Chinese Traditional ; Mice ; Principal Component Analysis ; Skin ; metabolism

OBJECTIVETo evaluate in vitro effect of abnormal savda munziq (ASMq) on the proliferation and apoptosis of human hypertrophic scar fibroblasts (HSFs).

METHODSHSFs were divided into six groups to receive different treatments as group A (blank control group), group B-E (ASMq in different concentration), and group F(5-Fu). Each group contains six specimens. The HSFs were cultured in vitro. After culture for 48 hours, the CCK8 test and flow cytometry methods were used to detect the proliferation, cell cycle and apoptosis.

RESULTSThe proliferation of HSFs in the B, C, D and E groups was inhibited at G2/M period, while it was inhibited at G0/S period in group F (P < 0.05). The inhibition effect of ASMq (0.1-1.0 mg/ml) on the fibroblasts enhanced in a concentration-dependent manner. Flow cytometry analysis with annexin V-FITC and PI staining confirmed the apoptotic. When HSFs were exposed to ASMq at 1.0 mg/ml (group E) for 48 h, the percentage of apoptotic cells increased to (43.7 +/- 2.58)%, which was significantly higher than that of blank control group (2.2 +/- 0.59)%. The induced apoptosis effect was also increased in a concentration-dependent manner.

CONCLUSIONASMq has a inhibitory effect on the proliferation and an enhancement effect on the apoptosis of fibroblast. ASMq could be used as an effective drug for treatment of hypertrophic scar.

Apoptosis ; Cell Cycle ; drug effects ; physiology ; Cell Division ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; pathology ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; In Vitro Techniques ; Medicine, East Asian Traditional

Objective To evaluate the association between methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism and depression risk.Methods Available studies containing genotype frequencies of MTHFR C677T were chosen,and pooled odds ratio (OR) with 95％ confidence interval (CI) was used to assess the strength of the association.Results Twenty-five eligible studies including 4 048 cases and 12 827 controls were identified.Meta-analysis showed a significant effect in the co-dominant model (T vs.C:OR 0 =1.25; Z=3.82,P=0.000;TTvs.CC:OR1=1.55,Z=3.79,P=0.000 ; except CT vs.CC:OR2=1.13; Z=1.78,P=0.076).Stratified analysis denoted that there was significantly higher risk in depression onset before 60 years old than depression onset after 60 years old.In subgroup analysis stratified by regressive meta-analysis,MTHFR C677T variant was statistically significantly relevant to depression risk in Chinese populations (OR 0 =1.58,Z=3.95,P=0.000; OR 1 =2.55,Z=3.92,P=0.000; OR 2 =1.43,Z=2.72,P=0.007),but not in no-Chinese populations (OR 0 =1.12,Z=2.10,P=0.035 ; OR 1 =1.23,Z=2.23,P=0.025 ; OR 2 =1.06,Z=0.74,P=0.456).Evidentiary strength rated by GRADE showed that 1 study moderate,3 studies low and 7 stdudies very low.Conclusion Our study provides evidence for a hereditary association between the MTHFR C677T variant and depression susceptibility,whereas there is an obvious variety in different regions,ethnicities and onset age.

BACKGROUND:celltherapy by the implantation of autologous bone marrow cells has been used for the treatment of ischemic heart diseases in clinical trials for decade. However, as the outcomes of celltransplantation obviously vary among patients, it is essential to identify the risk factors that may influence the level and function of progenitor cells in bone marrow, in order to identify the patients who would benefit the most from this treatment. OBJECTIVE:To observe the impact of perioperative cardiovascular risk factors on number and function of bone marrow progenitor cells from patients undergoing coronary artery bypass grafting surgery.
METHODS:We col ected clinical and laboratory data from 44 patients scheduled to undergo sternotomy for coronary artery bypass grafting procedures. Bone marrow was aspirated from the sternum during the operation and bone marrow mononuclear cells were isolated by density centrifugation with Ficol lymphoprep and then detected using trypan blue exclusion method. Levels of progenitor cells in bone marrow were evaluated using flow cytometry. Function of bone marrow progenitor cells were assessed by clonogenic and migration assays.
RESULTS AND CONCLUSION:We assessed the number of bone marrow mononuclear cells out of 20 mL bone marrow in duplicate samples from patients with coronary heart disease scheduled for coronary artery bypass grafting that was (10-89)×106 cells with over 95%activity. A negative correlation was observed between the number of bone marrow mononuclear cells and the age (n=44, r=-0.788, P=0.001). Levels of CD34+, CD133+, and CD34+CD133+cells in bone marrow mononuclear cells was (0.94±0.39)%, (0.46±0.28)%, and (0.53±0.26)%. Levels of CD34+cells and CD133+cells in patients with diabetes were significantly lower than those in patients without diabetes. Female, advanced age and poor heart function were related with reduced colony-forming ability of progenitor cells. A positive correlation was observed between level of CD34+cells and migration ability of bone marrow mononuclear cells. The results show that by density gradient centrifugation, we can harvest a sufficient number of bone marrow mononuclear cells in the treatment for ischemic heart disease. Age, gender, diabetes, heart function are correlated with bone marrow mononuclear cellnumber and functions.

Objective To observe any effect of low intensity pulsed ultrasound (LIPUS) on the expression of integrin-focal adhesion kinase (FAK)-mitogen-activated protein kinase (MAPK) mechanochemical transduction pathway-related proteins in the chondrocytes of rabbits with knee osteoarthritis (OA).Methods Of the 18 New Zealand white rabbits selected for the study,twelve received knee anterior cruciate ligament transection to model OA.The remaining 6 rabbits served as normal controls.At the 4th week after modeling the rabbits were sacrificed and chondrocytes were isolated and cultured in vitro.All the cultured cells were randomly divided into three groups:a normal control group (NC),an OA model group (OA) and an OA model plus LIPUS group (OA + LIPUS).When the cells had been cultured to the 2nd passage,the NC group and OA group cells had received no treatment.The OA + LIPUS group cells were exposed to 40 mW/cm2 of LIPUS for 20 min,once a day for 6 days.The expression of collagen protein type Ⅱ,aggrecan,MMP-13,integrin β1 p-FAK and p-p38,p-ERK1/2 and p-JNK Mapks were detected by Western blotting.Results Compared with the NC group,the expression of collagen type Ⅱ and aggrecan was significantly lower in the OA and OA + LIPUS groups,with more significantly lower expression of collagen type Ⅱ and aggrecan in the OA group than in the OA + LIPUS group.Compared with the NC group,the expression of MMP-13 was significantly higher in the OA and OA + LIPUS groups,with a significantly larger increase in the OA group.Compared with the NC group,the expression of integrin β1 and p-FAK was also significantly higher in the OA and OA + LIPUS groups,with a significantly larger increase in the OA + LIPUS group.Compared with the NC group,the expression of p-p38,p-ERK1/2 and p-JNK was also significantly higher in the OA group,but compared with the OA group,the expression of those kinases was,on average,significantly lower in the OA + LIPUS group.Conclusions LIPUS can inhibit the degradation of collagen type Ⅱ and aggrecan,and inhibit the expression of MMP-13,p-p38,p-ERK1/2 and p-JNK in OA chondrocytes,at least in vitro.At the same time,LIPUS can increase the expression of integrin β1 and p-FAK.The results show that LIPUS may activate an integrin-FAK-MAPK mechanochemical transduction pathway to induce changes in the extracellular matrix of chondrocytes.

Animals ; Lung ; immunology ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; physiopathology ; Spleen ; immunology ; pathology ; fas Receptor ; analysis

Objective To summarize the preliminary experience and early -stage follow -up results of simple transesophageal echocardiography(TEE)to guide right jugular internal vein for closure of atrial septal defect (ASD), and to investigate its feasibility.Methods A total of 24 ASD patients(12 male,12 female)treated by transcatheter ASD closure under solely guidance of TEE were summarized in Children Heart Center,People′s Hospital of Zhengzhou University from February 2015 to February 2016.Mean age,body weight and average diameter of the 24 cases of ASD were 5 -18 (12.4 ±5.6)years old,16 -60 (34.1 ±9.6)kg,5 -20 (11.0 ±2.5)mm,respectively.All patients were treated by right jugular internal vein transcatheter closure under solely guidance of TEE.The efficiency of imme-diate post -operation under TEE was estimated,and follow -ups were done at 24 hours,1 month,3,6 and 12 months after the procedures by transthoracic echocardiography,X -ray examination and electrocardiogram.Results A total of 23 patients were treated successfully under solely guidance of TEE.The diameter of devices was (14.5 ±3.6)mm.The procedural time was(20.5 ±7.2)min.One patient who had 2 ASD was treated by traditional thoracic surgery,because the small ASD had 5 mm residual shunt after the procedures.There were 3 cases with trivial residual shunt after the pro-cedures immediately.All patients could move at once after operation.The average follow -up time was(5.6 ±1.2) months.No patients suffered from residual shunt,occluder malposition,peripheral vascular injury,pericardial effusion or cardiac perforation during the follow -up.Conclusions For some selected patients,simple TEE guiding right jugular in-ternal vein for closure of ASD is applicable.The method not only prevents injury from radiation,but also shortens the in -bed time and avoid the lower limbs braking,with good early -stage follow -up results are desirable.

Objective:To investigate the expression and prognostic significance of CD19 in patients with Acute myelogenous leukemia with AML1-ETO positive. Methods: Clinical data of 66 patients AML with AML1-ETO positive who were newly diagnosed from Jan 2010 to Dec 2015 were collected. To retrospectively analyze the relationship between clinical characteristics and expression of CD19,so dose the prognosis. Results:The positive rate of CD19 expressing in AML with AML1-ETO positive was 50. 0%. There were no statistically significant differences in terms of age,gender,hemoglobin,platelet,percentage of bone marrow blasts,accompanied with chromosome ,gene mutations between patients with and without CD19 expression(P>0. 05). The white blood cell count(WBC) of the CD19 negative group was higher than CD19 positive group,while showed significant difference(P=0. 027). Although the relapse-free survival (RFS) of patients with CD19 expression was higher than those without,no significant difference was calculated (P=0. 105). Patients with CD19 expression had superior overall survival ( OS ) compared to those without CD19 expression ( P = 0. 030 ) . Multivariable analysis for OS identified CD19 positivity as an independent predictor associated with better prognosis. Conclusion: The expression of CD19 in AML with AML1-ETO positive may be an indicator associated with better prognosis.

Objective To study the morphological characteristics of femurs of adult human and 11 kinds of adult animals fromcattle, horses, pigs, goats, sheep, dogs, cats, rabbits, geese, ducks, chickens, and to establish an effective species identification method among various species. Methods The 4 cmmid-dia-physeal segment of the femur fromadult human (older than 20 years old) at autopsy w as obtained. Addi-tionally, the 4 cmones from11 kinds of adult animals w ere obtained. After decalcification, all femurs w ere made into slices, and then w ere observed by optical microscope. The 25 indexes w ere selected and analyzed by step discriminant analysis according to differences betw een human and mammal, human and poultry, and human and 11 kinds of animals. Results The histological structure of bone mineral density of middle part of femur had obvious characteristics among the species. And the morphology and number of osteon show ed the trend of obvious biological evolution. There w ere 11 indexes w ith significant differ-ences betw een human and 11 kinds of animals to establish some mathematical models to discriminate all species. The correct discrimination rate w as 96.3% betw een human and mammal. The correct discrimina-tion rate w as up to 100% betw een human and poultry, and w as 89.4% among human, mammal and poultry. Conclusion The mathematical models have good correct discrimination rate among human and the other animals, w hich could be applied in the practical species identification cases.