BACKGROUND:Many recent studies have shown a close relationship between inflammatory cytokines and osteoporosis and bone mineral density(BMD).However,the causal relationship between inflammatory cytokines and BMD has not been fully revealed. OBJECTIVE:To explore the potential causal relationship between inflammatory cytokines and BMD using a two-sample Mendelian randomization analysis. METHODS:The single nucleotide polymorphisms associated with 41 circulating inflammatory cytokines were selected from the open database of genome-wide association studies(GWAS)as instrumental variables.The GWAS data about BMD were from the Genetic Factors for Osteoporosis Consortium,involving a total of 32 735 individuals of European ancestry.Inverse variance weighting was used as the primary analysis to evaluate the causal effect.Weighted median,MR Egger regression,simple mode,and weighted mode methods were used to supplement the explanation.We used the MR-Egger intercept and MR-PRESSO method to conduct a pleiotropy test,the Cochran's Q test was used to determine whether there was heterogeneity in the results,and the leave-one-out method was used to evaluate the stability of the results.In addition,to more accurately assess the causality,the Bonferroni-corrected test was used to identify inflammatory cytokines that have a strong causal relationship with BMD. RESULTS AND CONCLUSION:(1)According to the results of the inverse variance weighting method,we found a positive causal relationship between interleukin-8 and lumbar spine BMD[β=0.075,95%confidence interval(CI):0.033-0.117,P=0.000 5),while a negative causal relationship between interleukin-17 and lumbar spine BMD(β=-0.083,95%CI:-0.152 to-0.014,P=0.018).There might be a negative causal relationship between tumor necrosis factor b and femoral neck BMD(β=-0.053,95%CI:-0.088 to-0.018,P=0.003),while a positive causal relationship between basic fibroblast growth factor and femoral neck BMD(β=0.085,95%CI:0.016-0.154,P=0.015).There might be a negative causal relationship between macrophage inflammatory protein-1a and total body BMD(β=-0.056,95%CI:-0.105 to-0.007,P=0.025).There was a negative causal relationship between interleukin-5(β=-0.019,95%CI:-0.031 to-0.006,P=0.004),stromal cell-derived factor-1a(β=-0.022,95%CI:-0.038 to-0.005,P=0.010),hepatocyte growth factor(β=-0.021,95%CI:-0.041 to-0.002,P=0.030),interleukin-4(β=-0.016,95%CI:-0.032 to-0.001,P=0.034)and heel BMD,while a positive causal relationship between nerve growth factor(β=0.019,95%CI:0.002-0.036,P=0.033),granulocyte colony-stimulating factor(β=0.011,95%CI:0.000-0.022,P=0.050),and heel BMD.Meanwhile,after the Bonferroni-corrected test,there was a strong positive causal effect between interleukin-8 and lumbar spine BMD(P=0.000 5).And consistent directional effects for all analyses were observed in MR Egger,weighted median,simple mode,and weighted mode methods.(2)Sensitivity analyses revealed no heterogeneity,pleiotropy,or outliers for the causal effect of circulating inflammatory cytokines on BMD.

Objective: To investigate the expression of lncRNA TUG1 (long non-coding RNA taurine up-regulated gene 1) in gastric cancer and its effect on the proliferation and apoptosis of gastric cancer cells. Methods: Surgically resected gastric cancer tissues and corresponding distal normal tissues (>5 cm away from the margin of tumor) of 40 gastric cancer patients from March 2016 to December 2017 at Ganzhou People's Hospital of Jiangxi Province were collected, and qPCR was used to examine the expression of lncRNA TUG1.AGS gastric cancer cells were transfected with lncRNATUG1 over-expression plasmids and siRNAs, and the effects of lncRNA TUG1 on cell proliferation and apoptosis were assessed by CCK-8, qPCR and Flow cytometry. Results: lncRNATUG1 expression was significantly increased in gastric cancer tissues as compared to normal tissues; and it was not correlated with gender, age, tumor size, infiltration depth of tumor, lymph node-metastasis, tumor differentiation and TNM staging. TUG1 over-expression significantly suppressed the expressions of CDKN1A, BAX and Caspase-3 in AGS gastric cancer cells, and decreased G1 phase proportion and apoptosis rate, but increased S phase proportion and cell viability; in contrast, TUG1 siRNA transfection significantly promoted the expressions of CDKN1A, BAX and Caspase-3, and increased G1 phase proportion and apoptosis rate, but decreased S phase proportion and cell viability. Conclusion: Up-regulated lncRNATUG1 promotes proliferation and inhibits apoptosis of gastric cancer cells.

To explore the effect of geniposidic acid (GPA) on the signal pathway of small heterodimer dimer receptor (SHP) and liver receptor homologue 1 (LRH-1) in cholestasis rats induced by alpha-naphthalene isothiocyanate (ANIT).
 Methods: Fifty SD rats were randomly divided into five groups: a blank group, an ANIT group, an ANIT+GPA (100 mg/kg) group, an ANIT+GPA (50 mg/kg) group, and an ANIT+GPA (25 mg/kg) group (n=10 in each group). The GPA were intragastrically given to rats for 10 days, and the control group and the ANIT group were given normal saline. At the eighth day of administration, all rats except the blank group were given 65 mg/kg ANIT once until the tenth day. After the last administration, serum total cholesterol (TC), triglyceride (TG) and total bile acids (TBA) were measured. The primary hepatocytes (RPH) were isolated from normal rats and cultured. The cells were divided into a blank group, an ANIT (40 μmol/L) group, an ANIT (40 μmol/L)+GPA (4.00 mmol/L) group (A4.00G group), an ANIT (40 μmol/L)+GPA (1.00 mmol/L) group (A1.00G group), and an ANIT (40 μmol/L)+GPA (0.25 mmol/L) group (A0.25G group). The mRNA transcription levels of SHP and cholesterol 7 alpha hydroxylase (CYP7A1) in RPH were detected by real-time-PCR, and the protein levels of SHP and CYP7a1 were detected by Western blotting. In the LRH-1 silence experiment, the RPH were divided into a blank group, a negative transfection group, a siRNA-LRH group (ZR group), a siRNA-LRH+GPA (4.00 mmol/L) group (ZR4.00G group), a siRNA-LRH+GPA (1.00 mmol/L) group (ZR1.00G group) and a siRNA-LRH+GPA (0.25 mmol/L) group (ZR0.25G group). The protein and mRNA levels of SHP, CYP7a1, LRH-1 were detected. In the over-expression experiment, the RPH were also divided into a blank group, a negative transfection group, a LRH-1 over-expression plasmid group (OE group), a LRH-1 over-expression plasmid+GPA (4.00 mmol/L) group (OE4.00G group), a LRH-1 over-expression plasmid+GPA (1.00 mmol/L) group (OE1.00G group), and a LRH-1 over-expression plasmid+GPA (0.25 mmol/L) group (OE0.25G group). The protein and mRNA levels of SHP, CYP7a1 and LRH-1 were detected.
 Results: Compared with the blank control group, TC and TBA were significantly increased (both P<0.01) in the ANIT group, but there was no difference in TG; compared with the ANIT group, the contents of TC and TBA in the AG100 and AG50 groups were significantly reduced (all P<0.01). Compared with the blank control group, the proteins and mRNA levels of SHP were significantly decreased (P<0.01), while CYP7a1 were dramatically increased (P<0.01) in the ANIT group; compared with the ANIT group, the proteins and mRNA levels of SHP in the A4.00G group and the A1.00G group were significantly increased (both P<0.01), while the levels of CYP7a1 proteins and mRNA levels were evidently decreased in the A4.00G and A1.00G groups (both P<0.01). Compared with the negative transfection group, the proteins and mRNA levels of CYP7a1 and LRH-1 were dramatically restrained (all P<0.01), while there was no change in SHP in the ZR group; compared with the ZR group, the proteins and mRNA levels of SHP were significantly increased (all P<0.01), while LRH-1 and CYP7a1 were not changed in the ZR4.00G, ZR1.00G and ZR0.25G groups. Compared with the negative transfection group, the protein and mRNA levels of CYP7a1 and LRH-1 were significantly suppressed in the OE group (all P<0.01). Compared with the OE group, the protein and mRNA levels of SHP were evidently increased in the OE4G and OE1G groups (all P<0.01), while LRH-1 and CYP7a1 were not changed in the OE4G, OE1G and OE0.25G groups.
 Conclusion: The over-expression of LRH-1 in RPH can up-regulate the mRNA and protein levels of CYP7a1. GPA can improve the biochemical and liver pathology of ANIT-induced cholestasis rats, which may be related to the decrease of CYP7a1 by activating SHP through LRH-1 in RPH.
Animals ; Cholestasis ; Iridoid Glucosides ; Rats ; Rats, Sprague-Dawley ; Receptors, Cytoplasmic and Nuclear ; Signal Transduction

Fast progresses in stem cell-based tooth tissue engineering have been achieved in recent years in several animal models including the mouse, rat, dog, and pig. Moreover, various postnatal mesenchymal stem cells of dental origin have been isolated and shown capable of differentiating into odontoblasts and generating dentin. Meanwhile, human keratinocyte stem/progenitor cells, gingival epithelial cells, and even iPSC-derived epithelium have been demonstrated to be able to differentiate into functional ameloblasts. Translational medicine studies in the nonhuman primate are irreplaceable steps towards clinical application of stem cell-based tissue engineering therapy. In the present study, we first examined the epithelial stem cell markers in the rhesus skin using immunostaining. Keratinocyte stem cells were then isolated from rhesus epidermis, cultured in vitro, and characterized by epithelial stem cell markers. Epithelial sheets of these cultured keratinocytes, which were recombined with E13.5 mouse dental mesenchyme that possesses odontogenic potential in the presence of exogenous FGF8, were induced to differentiate into enamel-secreting ameloblasts. Our results demonstrate that in the presence of appropriate odontogenic signals, rhesus keratinocytes can be induced to gain odontogenic competence and are capable of participating in odontogenesis, indicating that rhesus keratinocytes are an ideal epithelial cell source for further translational medicine study of tooth tissue engineering in nonhuman primates.
Ameloblasts* ; Animals ; Dentin ; Dogs ; Epidermis ; Epithelial Cells ; Epithelium ; Humans ; In Vitro Techniques ; Keratinocytes* ; Macaca mulatta ; Mental Competency ; Mesenchymal Stromal Cells ; Mesoderm* ; Mice* ; Models, Animal ; Odontoblasts ; Odontogenesis ; Primates ; Rats ; Skin ; Stem Cells ; Tissue Engineering ; Tooth ; Translational Medical Research

Objective To screen an ssDNA aptamer for rabbit mesenchymal stem cells (MSCs) and to identify the ability of the aptamer to recognize MSCs of a variety of species origin.Methods MSCs were isolated from the thigh bone of immature rabbits and identified by induced osteogenic and adipogenic differentiation,respectively.Aptamers were screened by cell SELEX (systematic evolution of ligands by exponential enrichment) technique targeting to isolated MSCs.Enrichment of the 5th pool was evaluated through binding assay of FAM modified pool to MSCs by confocal microscopy.The enriched 5th pool was then cloned into pGE-T vector and the cloned sequences were determined randomly.The candidates were chosen based on primary sequence conservation and predicted secondary structure by RNA structure and MEME online analysis.Flow cytometry analysis was used to identify the aptamers binding to MSCs of rabbit, rat, and human origin.Results The isolated MSCs had the potential of osteogenic differentiation and adipogenic differentiation under certain conditions.Aptamer 5-1-12 from 5th enriched pool was characterized as MSCs recognizing aptamer binding to MSCs of rabbit, rat and human origin.Conclusion Aptamer 5-1-12 that recognizes MSCs of different species origin is obtained through live cell-SELEX.

OBJECTIVE:To provide reference for improving the quality of pharmacy education in China.METHODS:Based on domestic and foreign related literatures in recent years,and the information from pharmacy school official websites of USA,to introduce to summarize the characteristics of the selection and admission methods and requriements of Doctor of Pharmacy students in pharmacy schools of universities in USA.RESULTS & CONCLUSIONS:The pharmacy education of USA developed rapidly in recent 10 years.To ensure the quality of pharmacist,the pharmacy schools attached great importance to selection and admission process of Doctor of Pharmacy students in USA.They did not only pay attention in academic scores,such as Pharmacy College Admission Test score(PCAT),Grade Point Average(GPA) in university before applying to pharmacy school,also used different ways to investigate others' evaluation and self-evaluation,the ability of the applicant to perform social activities by recommendation letters,personal statement and experience in extracurricular activity.Based on material evaluation and applicant comparison,according to proportion selection and applicant interview,oral and communication skills of the applicant were investigated.Finally,admission committee of pharmacy college makes decision.Compared with China,the pharmacy schools of American univerisities choose Doctor of Pharmacy students more comprehesively and pay more attention to the comprehensive ability beyond the applicant's score,which form a relatively complete system.

BACKGROUND:Conventional surgical repair can cause large traumas in patients with knee injuries, and patients often recover slowly after implant fixation, most of whom can appear to have poor recovery of knee function. OBJECTIVE: To explore the folow-up effect of arthroscopic double-steel wire clip fixation on tibial eminence avulsion fracture of anterior cruciate ligament. METHODS: A retrospective analysis was performed on the clinical data of 23 patients with tibial eminence avulsion fractures, who were given arthroscopic double-steel wire clip fixation. The patients were folowed up for 1-6 months. Short- and middle-term therapeutic effect as wel as IKDC and Lysholm scores before and after treatment were observed and analyzed. RESULTS AND CONCLUSION:The operation time was 35-65 minutes, and no complications, such as blood, nerve and anterior cruciate ligament injuries occurred. Moreover, no infection and other poor biocompatible reactions occurred after internation fixation. Al patients were folowed up for 1-6 months. The excelent and good rate was 87% at 1 month after treatment and 96% at 6 months after treatment. Al the patients had improved IKDC score and Lysholm score after treatment (P < 0.05), indicating that the knee function of patients was improved significantly.

Objective To observe the damage effect of borneol on rabbit corneal epithelial cells. Methods After the treatment with borneol at 100, 200, 400 μg·mL-1 respectively, the viability of rabbit corneal epithelial cells was determined by methyl thiazolyl tetrazolium ( MTT) assay, cell apoptosis was determined by flow cytometry with Annexin V- fluorescein isothiocyanate/propidium iodide ( FITC/PI) staining, and Caspase-3 mRNA expression was detected with real-time reverse transcription polymerase chain reaction ( RT-PCR) . Results Borneol at the concentrations of 100, 200, 400 μg·mL-1 inhibited the activity of rabbit corneal epithelial cells. Compared with the normal control group, borneol increased the rate of apoptosis, and enhanced the Caspase-3 mRNA expression in rabbit corneal epithelial cells ( P<0.05 or P<0.01) . Conclusion Borneol can inhibit the proliferation of rabbit corneal epithelial cells, induce cell apoptosis through enhancing the expression of apoptosis-related gene Caspase-3 mRNA.

OBJECTIVE:To study the bactericidal activity of thymopentin and its derived peptides. METHODS:Agar plate count was adopted to determine the bactericidal activity of thymopentin [arginine(R)-lysine(K)-aspartic acid(D)-valine(V)-tyro-sine(Y),RKDVY],its derived peptide 1 [RKN(agedoite,N)VY] and derived peptide 2(RKKVY)to Gram negative bacterial (Proteusbacillus vulgaris,Escherichia coli) and Gram positive bacterial (Staphylococcus aureus,Enterococcus faecium). There were 15.625-1 000 μg/ml for peptides,102 CFU for bacteria. RESULTS:Three pentapeptides possessed bactericidal activity against Gram negative bacteria. The activities of RKKVY and RKNVY were stronger than RKDVY(P<0.01),there was no significant dif-ference between RKKVY and RKNVY(P>0.05). They also possessed bactericidal activity against Gram positive bacteria,and the activity from strong to weak was RKKVY>RKNVY>RKDVY(P<0.01). CONCLUSIONS:Thymopentin and its derived peptides possess bactericidal activity against Gram negative and positive bacteria,with dose-effect relationship.

OBJECTIVETo select and identify ssDNA aptamers specific to Streptococcus mutans strains with different cariogenicity isolated from clinical specimens.

METHODSSubtractive SELEX technology targeting the whole intact cells was used to screen for ssDNA aptamers specific to the clinical isolates Streptococcus mutans strains with different cariogenicity. Radioactive isotope, flow cytometry, gene cloning and sequencing, MEME online software and RNA structure analysis software were employed to analyze the first and secondary structures of the aptamers and identify the screened aptamers.

RESULTSDetection by radioactive isotope showed sufficient pool enrichment after 9 rounds of subtractive SELEX. Flow cytometry showed that the selected aptamers H1, H16, H4, L1, L10 and H19 were capable of binding specifically with highly cariogenic Streptococcus mutans strains but not with strains with a low cariogenicity. The aptamer H19 had the strongest binding capacity to highly cariogenic Streptococcus mutans strains, with a dissociation constant of 69.45∓38.53 nmol/L.

CONCLUSIONWe have obtained the ssDNA aptamers specific to the clinical isolates of highly cariogenic Streptococcus mutans strains.

Aptamers, Nucleotide ; genetics ; Cloning, Molecular ; DNA Primers ; Dental Caries ; microbiology ; Gene Library ; Humans ; Nucleic Acid Conformation ; SELEX Aptamer Technique ; Species Specificity ; Streptococcus mutans ; classification ; genetics ; isolation & purification