Aim To investigate the effect of Ginsen-oside Rh2 on apoptosis in human colorectal cancer cell SW480,and to explore the possible mechanism of it. Methods The proliferation activity of SW480 treated with Ginsenoside Rh2 was measured CCK-8 assay.Ap-optosis rates were evaluated by FCM.Hoechst 33258 staining was used to observe cell nucleus morphologi-cal;change SW480 cells were treated with Ginsenoside Rh2,and the protein expressions of Bcl-2,Bax,p53, cleaved caspase-3 ,PI3 K,AKT,P-AKT,GSK-3β,P-GSK-3βwere detected by Western blot;SW480 cells were treated with LY294002,Rh2,LY294002+Rh2, the expressions of PI3 K,AKT,P-AKT,GSK-3β,P-GSK-3βwere detected by Western blot.Results The proliferation of SW480 cells was significantly inhibited by Ginsenoside Rh2 in dose-dependent and time-de-pendent manner.FCM showed the inducing apoptosis effect of Ginsenoside Rh2 was significantly different from that of control group.Hoechst 33258 staining in-dicated clearly cell apoptosis in Ginsenoside Rh2 treat-ment groups.Western blot showed Ginsenoside Rh2 decreased expression of Bcl-2,increased expression of Bax,p53 and cleaved caspase-3,PI3K/AKT/GSK-3βpathway proteins PI3 K,P-AKT,P-GSK-3βdecreased obviously,AKT and GSK-3βwere not changed signifi-cantly in SW480.SW480 cells were separately treated with LY294002,Rh2,LY294002 +Rh2,there were no significant difference in AKT and GSK-3βprotein a-mong all groups,and the expression of PI3 K,P-AKT, P-GSK-3βdecreased more obviously in LY294002 +Rh2 group compared with LY294002 and Rh2 alone. Conclusion Rh2 induces colorectal cancer cell apop-tosis through PI3 K/AKT/GSK-3βpathway,which ac-tivates p53 and cleaved caspase-3,and destroys the balance of Bcl-2/Bax.

Aim To investigate the effect of ribonucleic acidⅡon apoptosis in human leukemia cell lines K562 and KG1 a.Methods Cell counting kit-8(CCK-8)as-say was performed to detect proliferation activity of K562 and KG1 a cells treated with ribonucleic acidⅡ. Apoptosis index was assessed by flow cytometry(FCM) and fluorescent Hoechst 33258 staining was used for observing morphologic changes of apoptosis.Expres-sion levels of p53,Bax,Bcl-2 and cleaved caspase-3 were analyzed by Western blot.Results The prolifera-tion of K562 and KG1 a cells was significantly inhibited by ribonucleic acid Ⅱ treatment for 12 h,24 h,48 h at concentrations of 100~300 mg·L-1 ,which indica-ted the inhibitory effect of ribonucleic acid Ⅱ was in dose-dependent and time-dependent manners.FCM re-sults displayed a dose-dependent increase in cell apop-totic rate.Hoechst 33258 staining showed the typical apoptotic morphology in some leukemic cells treated with ribonucleic acid Ⅱ,including increased nuclear chromatin concentration and edge accumulation.West-ern blot analysis showed the increased expression of p53,Bax,cleaved caspase-3 and decreased expression of Bcl-2 in K562 and KG1 a cells treated with ribonu-cleic acid Ⅱ.Conclusions Ribonucleic acid Ⅱ can induce apoptosis of leukemia K562 and KG1 a cells by up-regulating p53,which mediates Bcl-2/Bax balance and activates caspase-3 .

OBJECTIVETo analyze the potential pathogenic genomic imbalance in children with unexplained intellectual disability (ID) and/or developmental delay (DD) and its association with phenotypes, and to investigate the value of array-based comparative genomic hybridization (array-CGH) in clinical molecular genetic diagnosis.

METHODSThe whole genome of 16 children with ID/DD was scanned by the array-CGH for detection of genomic copy number variations (CNVs), and the revealed genomic imbalance was confirmed by multiplex ligation-dependent probe amplification.

RESULTSG-band karyotyping of peripheral blood cells showed no abnormalities in the 16 children. The results of the array-CGH revealed that 6 (38%) of the 16 patients had genomic CNVs, and 3 cases of CNVs were normal polymorphic changes; 1 CNV was a microdeletion of 4p16.3, which was the critical region for Wolf-Hirschhorn syndrome, and 1 CNV was a microdeletion of 7q11.23, which was the critical region for Williams-Beuren syndrome. Moreover, a CNV was identified with two duplications at 2q22.2 and 15q21.3 in a boy, which proved to have a clinical significance due to its association with ID, brain DD, unusual facies, cryptorchidism, irregular dentition, etc.

CONCLUSIONSArray-CGH allows for the etiological diagnosis in some of the children with unexplained ID/DD. As a high-throughput and rapid tool, it has a great clinical significance in the etiological diagnosis of ID/DD.

Adolescent ; Child ; Comparative Genomic Hybridization ; methods ; Developmental Disabilities ; diagnosis ; genetics ; Female ; Humans ; Infant ; Intellectual Disability ; diagnosis ; genetics ; Male ; Multiplex Polymerase Chain Reaction

BACKGROUNDThe treatment for long bone defects has been a hot topic in the field of regenerative medicine. This study aimed to evaluate the therapeutic effects of calcium sulfate (CS) combined with platelet-rich plasma (PRP) on long bone defect restoration.

METHODSA radial bone defect model was constructed through an osteotomy using New Zealand rabbits. The rabbits were randomly divided into four groups (n = 10 in each group): a CS combined with PRP (CS-PRP) group, a CS group, a PRP group, and a positive (recombinant human bone morphogenetic protein-2) control group. PRP was prepared from autologous blood using a two-step centrifugation process. CS-PRP was obtained by mixing hemihydrate CS with PRP. Radiographs and histologic micrographs were generated. The percentage of bone regenerated bone area in each rabbit was calculated at 10 weeks. One-way analysis of variance was performed in this study.

RESULTSThe radiographs and histologic micrographs showed bone restoration in the CS-PRP and positive control groups, while nonunion was observed in the CS and PRP groups. The percentages of bone regenerated bone area in the CS-PRP (84.60 ± 2.87%) and positive control (52.21 ± 4.53%) groups were significantly greater than those in the CS group (12.34 ± 2.17%) and PRP group (16.52 ± 4.22%) (P < 0.001). In addition, the bone strength of CS-PRP group (43.10 ± 4.10%) was significantly greater than that of the CS group (20.10 ± 3.70%) or PRP group (25.10 ± 2.10%) (P < 0.001).

CONCLUSIONCS-PRP functions as an effective treatment for long bone defects through stimulating bone regeneration and enhancing new bone strength.

Animals ; Bone Regeneration ; drug effects ; Calcium Sulfate ; pharmacology ; Male ; Platelet-Rich Plasma ; Rabbits

Compartments of soil microorganism and enzymes between stereoscopic cultivation (three storeys) and field cultivation (CK) of Panax notoginseng were carried out, and the effects on P. notoginseng agronomic characters were also studied. Results show that concentration of soil microorganism of stereoscopic cultivation was lower than field cultivation; the activity of soil urea enzyme, saccharase and neutral phosphatase increased from lower storey to upper storey; the activity of soil urea enzyme and saccharase of lower and upper storeys were significantly lower than CK; agronomic characters of stereoscopic cultivated P. notoginsengin were inferior to field cultivation, the middle storey with the best agronomic characters among the three storeys. The correlation analysis showed that fungi, actinomycetes and neutral phosphatase were significantly correlated with P. notoginseng agronomic characters; concentration of soil fungi and bacteria were significantly correlated with the soil relative water content; actinomycete and neutral phosphatase were significantly correlated with soil pH and relative water content, respectively; the activities of soil urea enzyme and saccharase were significantly correlated with the soil daily maximum temperature difference. Inconclusion, The current research shows that the imbalance of soil microorganism and the acutely changing of soil enzyme activity were the main reasons that caused the agronomic characters of stereoscopic cultivated P. notoginseng were worse than field cultivation. Thus improves the concentration of soil microorganism and enzyme activity near to field soil by improving the structure of stereoscopic cultivation is very important. And it was the direction which we are endeavoring that built better soil ecological environment for P. notoginseng of stereoscopic cultivation.
Hydrogen-Ion Concentration ; Panax notoginseng ; growth & development ; Phosphoric Monoester Hydrolases ; metabolism ; Soil ; chemistry ; Soil Microbiology ; beta-Fructofuranosidase ; metabolism

AIM:To study the influence of bone marrow mesenchymol stem cell-drived exosomes(BMSC-exo-somes)on hindlimb activity,and the numbers of reactive astrocytes and residual neurons in spinal cord injury(SCI)rats. METHODS:BMSCs were cultured using the whole bone marrow adherent culture method and surface markers CD 90 and CD34 were verified by flow cytometry.Exosomes were isolated by ultracentrifugation and the morphology of exosomes was observed under transmission electron microscope.The protein markers CD63 and CD9 were verified by Western blot.After exosomes were applied to SCI rats,the Basso,Beattie and Bresnahan locomotor rating scale score,the Nissl staining of the lesion site,and the numbers of reactive astrocytes and residual neurons were assessed at various time points.RESULTS:Transmission electron microscopic observation revealed the presence of saucer -shaped vesicles.BMSC-exosomes were found to express high levels of CD63 and CD9.Compared with injury group,significant improvement of hindlimb activity scores from day 14 after injury in treatment group was observed(P<0.05),and less reactive astrocytes and more residual neu-rons from day 7 after injury were also observed(P<0.05).CONCLUSION:BMSC-exosomes inhibit reactive astrocytes and death of neurons,and improve hindlimb activity in the rats after SCI.

A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood. The technology utilizes special primers and Taqman MGB fluorescence probe to measure amplification products from the gag-pro-pol polyprotein gene of HTLV-I. HTLV-I copy number was normalized to the amount of cellular DNA by quantitation of the beta-actin gene, The amplification system was sensitive to detect 5 copy/microL. The standard curve had a good linearity when the quantity for the gene was between 10(3) and 10(7) copy/microL (R2 = 0.999). Good reproducibility was observed in each intra- and inter-assay. We also measured proviral load in peripheral blood in 12 HTLV-I seropositive former blood donors. Proviral load for HTLV-I infected donors ranged from 0.015 to 12.819 copy/cell in WBC with the mean of 3.116 copy/cell.
Gene Products, gag ; genetics ; Gene Products, pol ; genetics ; Human T-lymphotropic virus 1 ; genetics ; isolation & purification ; Humans ; Molecular Probes ; Polymerase Chain Reaction ; methods ; Viral Proteins ; genetics

OBJECTIVETo verify the regulation of acyl-coenzyme A:cholesterol acyltransferase 2 (ACAT 2), which is associated with cholesterol metabolism, by saturated fatty acids (SFAs).

METHODSPalmitic acid (PA), the most abundant saturated fatty acid in plasma, and oleic acid (OA), a widely distributed unsaturated fatty acid, were used to treat hepatic cells HepG2, HuH7, and mouse primary hepatocytes. In addition, PA at different concentrations and PA treatment at different durations were applied in HepG2 cells. In in vivo experiment, three-month male C57/BL6 mice were fed with control diet and SFA diet containing hydrogenated coconut oil rich of SFAs. The mRNA level of ACAT2 in those hepatic cells and the mouse livers was detected with real-time polymerase chain reaction (PCR).

RESULTSIn the three types of hepatic cells treated with PA, that SFA induced significant increase of ACAT2 expression (Pü0.01), whereas treatment with OA showed no significant effect. That effect of PA was noticed gradually rising along with the increase of PA concentration and the extension of PA treatment duration (both Pü0.05). SFA diet feeding in mice resulted in a short-term and transient increase of ACAT2 expression in vivo, with a peak level appearing in the mice fed with SFA diet for two days (Pü0.05).

CONCLUSIONSFA may regulate ACAT2 expression in human and mouse hepatic cells and in mouse livers.

Animals ; Base Sequence ; Cell Line, Tumor ; DNA Primers ; Dose-Response Relationship, Drug ; Fatty Acids ; pharmacology ; Humans ; Liver ; enzymology ; Male ; Mice ; Mice, Inbred C57BL ; Sterol O-Acyltransferase ; metabolism

BACKGROUNDAlthough great advances in techniques for noninvasive prenatal diagnosis using fetal cells from maternal peripheral blood have achieved, current technology does not meet the demands required for clinical use. In this study, we aimed to establish reliable methods for the gene analysis of fetal cells from maternal peripheral blood.

METHODSPrimed extension preamplification (PEP)-polymerase chain reaction (PCR), multiple primed in situ labeling (PRINS), and nested PCR were individually applied to detect the sex determining region Y (SRY) gene in single fetal cells collected from maternal peripheral blood.

RESULTSThe sensitivity and specificity of the detection of the SRY gene by PEP-PCR were 97.39% (149/153) and 99.17% (119/120), respectively. The sensitivity and specificity of PRINS were 97.56% (40/41) and 100% (35/35), respectively. The sensitivity and specificity of nested-PCR were 80.00% (24/30) and 87.50% (14/16), respectively.

CONCLUSIONSPEP-PCR and PRINS are reliable techniques for the gene analysis of single fetal cells from maternal peripheral blood because of their high sensitivity and specificity. PEP-PCR and PRINS can be used as standard methods of noninvasive prenatal diagnosis using single fetal cells from maternal peripheral blood.

Adult ; Female ; Genes, sry ; Humans ; Polymerase Chain Reaction ; methods ; Pregnancy ; blood ; Prenatal Diagnosis ; methods ; Sensitivity and Specificity

AIMTo promote the provision of reproductive health services to young people by exploring the attitudes and perceptions of university students in Shanghai, China, toward reproductive health.

METHODSFrom July 2004 to May 2006, 5 243 students from 14 universities in Shanghai took part in our survey. Topics covered the demands of reproductive health-care services, attitudes towards and experience with sex, exposure to pornographic material, and knowledge on sexual health and sexually transmitted infections (STIs)/AIDS.

RESULTSOf the 5 067 students who provided valid answer sheets, 50.05% were female and 49.95% were male, 14.86% were medical students, and 85.14% had non-medical backgrounds. A total of 38.4% of respondents had received reproductive health education previously. The majority of students supported school-based reproductive health education, and also acquired information about sex predominantly from books, schoolmates, and the Internet. Premarital sexual behavior was opposed by 17.7% of survey participants, and 37.5% could identify all the three types of STIs listed in the questionnaire. Although 83.7% knew how HIV is transmitted, only 55.7% knew when to use a condom and 57.8% knew that the use of condoms could reduce the risk of HIV infection.

CONCLUSIONThe reproductive health service is lagging behind current attitudes and demands of university students. Although students' attitudes towards sexual matters are liberal, their knowledge about reproductive health and STIs/AIDS is still limited. It is therefore necessary to provide effective and confidential reproductive health services to young people.

Adolescent ; Adult ; Attitude to Health ; China ; Female ; HIV Infections ; prevention & control ; transmission ; Health Knowledge, Attitudes, Practice ; Health Services Needs and Demand ; statistics & numerical data ; Health Surveys ; Humans ; Male ; Perception ; Reproductive Health Services ; utilization ; Sexual Behavior ; Sexually Transmitted Diseases ; prevention & control ; transmission ; Student Health Services ; utilization ; Surveys and Questionnaires ; Universities