Objective Genotype data of nine CODIS STR loci were gathered to examine the features of population differentiation and gene flow of seven Xinjiang minorities.Methods Heterozygosity,Nei's coefficient of genetic differentiation,Nei's genetic distance and Wright's F-statistics were calculated. Statistical tests using exact method were performed to measure the level of differentiation.Phylogenetic trees were constructed by Mega;AMOVA was processed by Arlequin.R-matrix model had been applied to describe the patterns of gene flow.Results It shows that average genetic heterogeneity for each population was above 0.7 with genetic differentiation coefficient below 2%.Statistical tests for population differentiation were significant for most of the loci.Phylogenetic analysis and AMOVA showed that all populations were divided into three main groups.The R-matrix analysis reflected that Uygur,Kirgiz and Ozbek had more amounts of gene flow than other populations,while the pattern of Hui was more isolated.Conclusion The seven minorities in Xinjiang are independent populations,while the level of differentiation is at average.The relationship in evolution is not far from each other,with wide gene flow.

Objective·To investigate the effects of interleukin-1β (IL-1β) on neonatal rat alveolar arrest induced by intra-amniotic injection of lipopolysaccharide (LPS). Methods·A neonatal SD rat model of bronchopulmonary dysplasia (BPD) was constructed by intra-amniotic injection of LPS in pregnant rats. The pregnant rats (E19) were randomly assigned to Saline group, LPS group and LPS+anti-IL-1β group. The lungs of the neonatal rats were randomly collected 1, 3 and 7 days after birth. Pathological changes in the lungs were observed by hematoxylin-eosin (H-E) staining, and expression of IL-1β mRNA and protein was detected by real-time PCR and ELISA, respectively. Rat bone marrow derived primary macrophage was cultured in vitro, and given LPS intervention, then genes related with IL-1β were detected through whole transcriptome sequencing. Results·Compared with the Saline group, the alveolar counts and secondary septa counts significantly decreased, and mean liner intercept significantly increased in LPS group. Moreover, the expression of IL-1β mRNA and protein in lungs significantly increased in LPS group. The LPS-induced pathological changes of lung tissues in neonatal rats were improved by anti-IL-1β. LPS could up-regulate the expression of genes including Gbp5, Ccl3, Nod2, Ccr5, Mefv, Casp4 and Ifnar1, but down-regulate Lgals9 and Gstp1. Among these genes Gbp5, Ccl3, Nod2, Ccr5, Casp4, Ifnar1 and Lgals9 could positively regulate IL-1βproduction. Conclusion·LPS can induce alveolar arrest through up-regulating the expression of IL-1β in macrophages in neonatal rat BPD model. Whole transcriptome sequencing reveals that LPS can regulate the expression of IL-1β in macrophages through several paths.

Objective This paper was to study the absorption and metabolism differences of intestine and liver for multicomponent licorice.Methods The components were identified with the UPLC-MS/MS. In situ closed-loop method was used to carry out the comparative experiments of absorption and metabolism differences between intestine and liver.Results 13 components were identified by UPLC-MS/MS. The absorption and metabolism results indicated some components in licorice water extract could be absorbed into blood and metabolism happened during this process. 14 metabolites were detected in the plasma sample. The hepatic metabolism results indicated many components could experience complex metabolism and more metabolites could be generated.Conclusions Liver was the major metabolism organ for licorice water extract and some components could be metabolized along with the absorption process in intestine. The absorption and metabolism differences between intestine and liver were significant.

Objective:To obtain the prokaryotic expression product for the group 6 allergen of Dermatophagoides farine ( Der f 6) and detect its IgE-binding rates with sera from asthmatic children. Methods: By enzyme digestion of pET28a (+)-Der f 6 with BamHⅠ plus XhoⅠ,the target gene Der f 6 was obtained and linked into the vector pET32a (+) to construct the recombinant plasmid pET32a(+)-Der f 6, which was then transfected into E. coli BL21 cells for expression, induced with isopropyl-β-D-thiogalactoside ( IPTG) ,purified by affinity chromatography and identified by SDS-PAGE,Western blot and AMLDI-TOF,and tested by ELISA for IgE reactivity with sera from asthmatic children. Results:The plasmids pET32a(+)-Der f 6 were constructed,transformed into E. coli BL21 and expressed successfully. SDS-PAGE of the purification product showed a specific band,Western blot showed the successful binding between the purification product and the His-tag in the plasmids,and MALDI-TOF/TOF identified the identical structure to the allergen Der f 6. Using the ELISA method developed with the recombinant proteins as coating antigen,the positive rate was 41. 3% (19/46) in asthmatic children allergic to dust mite. Conclusion: The plasmids pET32a (+)-Der f 6 were constructed successfully,expressed in E. coli BL 21 (DE3). The recombinant fusion protein has a good reactivity with sera from asthmatic children.

Objective To evaluate the role of autophagy in the development of inflammatory pain in the rats.Methods Twenty-four healthy adult male Sprague-Dawley rats,weighing 180-240 g,were randomly divided into 4 groups (n=6 each) by using a random number table:control group (group C),inflammatory pain group (group IP),dimethyl sulfoxide (DMSO) group (group D),and rapamycin (autophagy inducer) group (group R).Inflammatory pain was produced by injecting 50 μl bee venom into the plantar surface of the left hindpaw.In group C,0.9％ normal saline was injected into the plantar surface of the left hindpaw.In group D,2％ DMSO was injected through a gastric tube into the stomach 1 ml per day for 3 consecutive days,and the model was established at 1 h after injection on 3rd day.In group R,rapamycin l0 mg/kg (in 2％ DMSO) was injected through a gastric tube into stomach 1 ml per day for 3 consecutive days,and the model was established at 1 h after injection on 3rd day.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 2 h after the model was established.After measurement of the pain threshold,the dorsal horn of the spinal cord was removed for determination of the expression of microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ),Beclin-1 and p62 by Western blot.Results Compared with group C,the MWT was significantly decreased,the TWL was significantly shortened in IP and D groups,and the expression of LC3 Ⅱ,Beclin-1 and p62 in the dorsal horn of the spinal cord was significantly up-regulated in IP,D and R groups (P＜0.05 or 0.01).Compared with group IP,the MWT was significantly increased,the TWL was significantly prolonged,the expression of LC3 Ⅱ and Beclin-1 in the dorsal horn of the spinal cord was significantly up-regulated,and the expression of p62 was significantly down-regulated in group R (P＜0.05),and no significant change was found in the parameters mentioned above in group D (P＞0.05).Conclusion Autophagy disorders are involved in the development of inflammatory pain in the rats.

Objective To establish a high purity primary culture methods of human nasal epithelial cells (HNEC) in vitro and to provide a successful primary culture model for evaluation experiments of the nasal preparation .Methods Primary culture of human nasal epithelial cells were performed with enzymatic dissociation of isolated tissue and cultured in serum-free medium .HNEC were separated through magnetic field by immunomagnetic beads .We determined the purity of the separated cells by light microscopy and flow cytometry .The morphology of HNEC was observed with a scanning electron microscope .Results Under an inverted phase microscope ,the cells morphology was paving stone shaped .Under the scanning electron microscopy ,abundant microvilli and cilia differentiation were observed .Flow cytometry showed the epithelial cells accounted for 99% .Conclusion The highly purified HNEC can be directly isolated by the magnetic cell sorting system .The cell model can be used for the basic research of nasal cavity preparation .

Objective:To compare structure and function of mite Der f 3,Der p 3 and Eur m 3.Methods: Obtained mite allergens amino acid sequences from the International Union of Immunological Societies nomenclature database .Then physiochemical characterization,sequence alignment,secondary structure,three dimensional (3D) structure and epitopes of three proteins were analyzed by bioinformatics methods.Results:According to results of alignment ,Der f 3,Der p 3 and Eur m 3 displayed 88.51%sequence iden-tity.Der f 3,Der p 3 and Eur m 3 all contained three active sites and two trypsin functional domains ,which showed high identity of amino acid residues.Active sites of three proteins ,which closing to each other in three dimensional (3D) structure,constituting the active center of the enzyme.Secondary and 3D structure of three proteins all contains α-helices,β-sheets and random coils.Epitopes analysis revealed that Der f 3,Der p 3 and Eur m 3 all have 5 main potential epitopes located in random coils.Epitope sequences of Der f 3,Der p 3 and Eur m 3 overlapping in three domains (peptides of 79-81aa,129-135aa and 172-174aa),but the residues in these three domains were not identical.Conclusion:These studies lay the foundation for biochemical and genetic analysis of these 3 allergens,and may contribute to vaccine development for allergen-specific immunotherapy.

ObjectiveComparingfourgroupsofimmunosuppressivemousemodelsestablishedby cyclophosphamide administration in different doses and periods , we used partial least squares ( PLS) analysis to evaluate the immunosuppressive mouse models comprehensively and select the appropriate way to establish this model .Methods In this experimental study, 58 male KM mice were randomly divided into five groups: normal group (10 mice) were given normal saline daily by i.p.injection, model group 1 (12 mice) was given 40 mg/kg CTX daily by i.p.injection for 10 days, model group 2 (12 mice) was given 80 mg/kg CTX daily by i.p.injection for 3 days, model group 3 (12 mice) was given 40 mg/kg CTX daily by i.p.injection twice for a week, model group 4 (12 mice) was given 50 mg/kg CTX daily by i.p.injection for 7 days.After the injection of cyclophosphamide , the daily metabolic activities were detected everyday such as body weight , water intake , and food intake , organ index and immunological indexes such as blood RT of the model mice were measured as well .Using partial least squares ( PLS) analysis to the models and analyzing the final weight , final anal temperature , organ index , and blood routine examination in order to evaluate the immunosuppressive mouse models comprehensively .Results Compared with the normal group , different dosages of CTX reduced the weight and anal temperature of model mice (P<0.05), the food intake and water intake (P<0.01), and the spleen index and thymus index ( P<0.01 ) .Besides , the amount and percentage of basophilic granulocytes decreased ( P <0.05 ) , and the percentage of MCHC , macrophage went up , as well as the absolute value of WBC and lymphocytes were decreased in the model groups 1, 2, and 4 (P<0.05).According to the PLS analysis, there were significant differences among models 1, 2, and 4 when compared with the normal group , especially the most different in the model group 1.Conclusions After the PLS analysis of different indexes , the optimal way to establish immunosuppressive mouse models is the procedure with 40 mg/kg CTX daily injected i .p.for ten days .Our findings provide experimental evidence for the establishment of immunosuppressive mouse models .

Objective:To investigate the effect of the extract of Schisandra chinensis on the matrix metalloproteinases(MMPs) system in kidney tissue of the diabetic rats,and to explore its protective effect on the kidney tissue from the matrix degradation perspective.Methods:STZ was used to establish rat models of diabetes mellitus.A total of 45 diabetic rats were randomly divided into model group,extract of Schisandra chinensis group and Benazepril group,and there were 15 rats in each group.Another 15 rats were selected and used as normal control group.12 weeks after administration,the routine blood and urine biochemical indexes,the histological changes,blood glucose (BG),blood urea nitrogen(BUN),serum creatinine(Scr),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterin(LDL-C),total cholesterol(T-CHO),and triglyceride(TG) levels,excretion rates of albuminuria and proteinuria of the rats in various groups were detected;the expression amounts of fibronectin (FN),type Ⅳ collagen (Col Ⅳ),and matrix metalloproteinase inhibitor metalloproteinase-2 (TIMP-2) in kidney tissue of the rats were detected by immunohistochemical method;the activity of matrix metalloproteinase-2 (MMP-2) was detected by zymography.Results:Compared with model group,the glomeruli matrix accumulation of the rats in extract of Schisandra chinensis group was significantly improved,the excretion rate of albuminuria,LDL-C level and serum MDA level were decreased(P<0.05),the activities of CAT(P<0.01)and SOD(P<0.05)in kidney tissue were increased,and the level of MDA in kidney tissue was decreased(P<0.05).The immunohistochemistry results showed that compared with model group,the expression amounts of FN,Col Ⅳ,and TIMP-2 in kidney tissue of the rats in extract of Schisandra chinensis group were significantly decreased.The zymography results showed that compared with model group,the activity of MMP-2 in kidney tissue of the rats in extract of Schisandra chinensis group was significantly increased(P<0.05).Conclusion:Extract of Schisandra chinensis has protective effect on the kidney tissue of the diabetic rats induced by STZ,and the mechanism may be related to the inhibition of oxidative stress and the improvement of MMP-2 activity as well as the inhibition of TIMP-2 expression which could improve the matrix degradation.

This study was aimed to determine the solubility an d oil-water partition coefficient of main active com-ponents in Ge-Gen Qin-Lian (GGQL) Tablets (puerarin, baicalin and berberine hydrochloride) in phosphate buffer solution of different pH values and under the background of many components. Solubility of puerarin, baicalin and berberine hydrochloride in different medium pH, and oil-water partition coefficient of the octanol-water and oc-tanol-buffer system were determined by HPLC method. The results showed that the solubility and oil-water partition coefficient of puerarin, baicalin and berberine hydrochloride were varied with the change of pH, and varied under the background of components. At pH 7.4, the solubility was the biggest;puerarin was 7.56 mg·mL-1;baicalin was 17.07 mg·mL-1; berberine hydrochloride was 3.57 mg·mL-1. Oil-water partition coefficient P of these components at pH 1.0 was bigger;puerarin was 0.420 (lgP=-0.38);baicalin was 10.783 (lgP=1.03);berberine hydrochloride was 0.267 (lgP=-0.57). It was concluded that lipid solubility of puerarin, baicalin and berberine hydrochloride at pH 1.0 was better. It was speculated that better absorption in the stomach, and low lipid solubility under other pH. It was speculated that lipid solubility may be one of the reasons affecting the intestinal absorption.