Objective To investigate the effect of insulin-like growth factor- Ⅰ ( IGF- Ⅰ ) on the proliferation and osteogenesis of human periodontal ligament stem cells ( hPDLC ) under three-dimensional (3D) culture system. Methods Human periodontal cells were isolated from the ligament of surgically extracted human teeth, and through the limiting dilution assay, got mono-clone of the cell, hPDLCs were isolated from MesenPRO RS medium. Rotary cell culture system (RCCS) was enrolled to set 3D environment. Control group and experiment groups were assigned according to the concentration of IGF- Ⅰ .There were 5 level of experiment groups (0.1,1,10,50,100 μg/L). Proliferation was tested with methyl thiazolyl tetrazolium ( MTT), and alkine phosphatase (ALP) level was assayed by spectrophotometer to analyze the osteogenesis of hPDLCs. Gene expression of ostetocalcin(OCN)and type Ⅰ collagen (Col Ⅰ )were assayed by reverse transcriptase polymerase chain reaction(RT-PCR). Results In 3D culture system,the effect of IGF- Ⅰ on cell proliferation was significantly different between control group and experiment groups( P ＜ 0.05 ), and there showed significant differences between the group of 0.1 μg/L ( 0.219 ±0.021 ) IGF- Ⅰ and the groups of 50, 100 μg/L(0.287 ±0.011,0.293 ±0.012). However, there showed no significant differences among other groups. Significant differences of ALP activity were observed between the control group and experiment groups, and between the groups of 1, 10 μg/L(0.304 ±0.020, 0.310 ±0.013) and that of 50, 100 μg/L (0.347 ±0.011, 0.344 ±0.010) (P ＜0.05). While no significantdifferences were detected between the group of 1 μg/L and that of 10 μg/L, nor between the group of 50 μg/L and that of 100 μg/L. Expressions of Col Ⅰ and OCN in mRNA and protein level both showed dose-dependent increase. Conclusions In 3D culture system, in the scale of 0.1-100 μg/L, the effect of IGF-Ⅰ on the proliferation of hPDLCs increased dose-dependently. 100 μg/L IGF- Ⅰ promotes osteogenesis of the cells significantly.

OBJECTIVETo investigate the effect of insulin-like growth factor- I (IGF- I) on the proliferation and alkaline phosphatase (ALP) activity of human periodontal ligament cells (hPDLCs) under three-dimensional (3D) culture system.

METHODSThe hPDLCs were cultured from periodontium of human teeth by the outgrowth method. Rotary cell culture system (RCCS) was enrolled to set 3D culture system. Samples were set to four groups: Negative control group, positive control group (3D group, IGF-I group), and experimental group (3D with IGF- I group). Proliferation was tested with methylthiazolyl tetrazolium (MTT), and ALP activity was assayed by spectrophotometer at 1, 3, 5, 7 d respectively.

RESULTSCompared with that of negative control group, cell proliferation increased significantly in 3D with IGF-I group since 3 d (P < 0.05). Besides, the cell proliferation of 3D with IGF-I group was significantly higher than that of 3D group (P < 0.05). ALP activity of 3D with IGF- I group was significantly higher than that of negative control group, and 3D group at 3, 5, 7 d (P < 0.05).

CONCLUSIONIGF-I significantly promotes the proliferation and ALP activity of hPDLCs under 3D culture system.

Alkaline Phosphatase ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Humans ; Insulin-Like Growth Factor I ; Periodontal Ligament ; Somatomedins

OBJECTIVETo observe the change of lymphatic reactivity to substance P (SP) during the process of hemorrhagic shock (HS) with a technique of lymphatic perfusion in vitro in this study.

METHODSMale Wistar rats were randomly divided into control group (surgical procedure only) and HS group (the rats in this group were further divided into five subgroups: shock 0 h, 0.5 h, 1 h, 2 h and 3 h groups after duplicating the HS model with method of bloodletting to mean arterial blood pressure was 40 mmHg through the femoral venous). Thoracic ducts were separated from HS rats at the corresponding time points in each group. A segment of thoracic duct was pressed and perfused in vitro at transmural pressure of 3 cm H2O, and then stimulated with gradient SP respectively. The end systolic diameter, end diastolic diameter, contraction frequency (CF) and passive diameter of isolated lymphatics were measured, while the contraction amplitude (CA), tonic index (TI) and fractional pump flow (FPF) were calculated, and the different values between pre- and post- administration of SP of CF, CA, TI and FPF were calculated and expressed as Delta CF, Delta TI, Delta CA and Delta FPF to further assess the reactivity of lymphatics.

RESULTSAfter SP incubation, the Delta CF, Delta TI, Delta CA and Delta FPF of 0 h- and 0.5 h shocked lymphatics were significantly increased when compared with that of control group on one or several concentrations. The Delta CF (at 3 x 10(-7) mol/L of SP) and Delta TI (1 x 10(-7) mol/L) of 2 h- shocked lymphatics and the Delta CF (1 x 10(-7) mol/L, 3 x 10(-7) mol/L), Delta TI (1 x 10(-7) mol/L) and Delta CA (1 x 10(-7) mol/L) of 3 h- shocked lymphatics were all significantly reduced when compared with control group.

CONCLUSIONThe reactivity of lymphatics to SP presented a biphasic change during the process of HS: increase in early phase and decline in later stage.

Animals ; Lymphatic Vessels ; physiopathology ; Male ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; physiopathology ; Substance P ; analysis ; Thoracic Duct ; physiopathology

Objective To construct an over-expressing plasmid containing full length of PELP1 gene and investigate the characterization of PELP1 expressions in different tissues.Methods Full length PELP1 gene was cloned using RT-PCR product of MCF-7 total RNA as the template.The PELP1 gene fragament was amplified by PCR and two enzyme sites EcoR I and Xho I were added.Then PELP1 gene was cloned into the pIRES2-EGFP palsimd (containing EcoR I and Xho I enzyme sites) to make reconstructed plasmid pIRES2-EGFP-PELP1.Enzyme digestion and sequencing were employed to assess the integrity and correctness of PELP1 gene cloned.The plasmid pIRES2-EGFP-PELP1 was transfected into human periodontal ligament stem cells,and the expression of PELP1 was examined by quantitative real-time-PCR or Western-blotting.Results The integrity and correctness of PELP1 gene were confirmed by digestion and sequencing.Conclusions The full length of PELP1 gene from MCF-7 cell was successfully cloned and over-expression plasmid pIRES2-EGFP-PELP1 constructed.

OBJECTIVETo summarize the clinical and ultrasonic features of breast cancer in women aged 80 and older.

METHODSA total of 60 patients (62 lesions) aged 80 and older with pathologically confirmed breast cancer from September 1993 to October 2012 were enrolled in this study and their clinical manifestations, ultrasonic features, therapeutic methods, and prognoses were analyzed.

RESULTSMost patients (83.3%) went to see a doctor because of nodules touched by themselves. The average diameter of the carcinoma was (2.4±1.1)cm. Most tumors (75.8%) were invasive ductal carcinomas, followed by the mucinous carcinoma (11.3%). Among the 45 lesions with ultrasound records, 40 (88.9%) were irregular in morphology; the aspect ratio of 35 lesions (77.8%) was less than 1;24 lesions (53.3%) had indistinct boundary;calcification existed in 21 lesions (46.7%); and 16 lesions (35.6%) had rear echo attenuation. The preoperative diagnostic accuracy of ultrasonography was 93.5%. In addition, 45 patients (75.0%) underwent breast tumor extended resection, 13 (21.7%) received modified radical mastectomy, 2 patients (3.3%) underwent simple breast resection. No death was noted during the operation and there was no major peri-operative complications. Of 31 patients with complete follow-up records, 7 had recurrence or metastasis and 1 died of heart disease.

CONCLUSIONSMost breast cancers in women older than 80 years are relatively large, with typical ultrasonic features. The preoperative diagnosis is often accurate. Few lymphatic metastases exist, and the prognosis is good. Conservative surgeries are preferred for these elderly patients.

Aged, 80 and over ; Breast Neoplasms ; diagnostic imaging ; surgery ; Carcinoma ; surgery ; ultrastructure ; Female ; Humans ; Prognosis ; Ultrasonography

In order to construct a pichia pastoris expression vector containing the extracellular domain of human Jagged1 and the Fc fragment of human IgG1 fusion gene, or containing only the Fc fragment of human IgG1 and to express them in pichia pastoris. The extracellular domain of human Jagged1 gene was cloned from normal human bone marrow cells. After DNA sequencing, the extracellular domain of Jagged1 gene was inserted into pIC-Fc vector constructed previously, which is Pichia pastoris expression vector containing only the Fc fragment of human IgG1. The constructed plasmid was transformed into yeast GS115 by means of electroporation. The recombinant transformants with a high copy number of the plasmid were selected on MD plate with G418. The expression of protein was induced by addition of methanol. Then, protein expression was analyzed by SDS-PAGE. The results indicated that the extracellular domain of human Jagged1 gene was effectively amplified. The DNA sequencing result showed that the constructed plasmid containing hJagged1(ext)-Fc fusion gene was the same as designed. The fusion protein was successfully expressed in Pichia pastoris. It is concluded that the hJagged1(ext) gene has been successfully cloned and expressed, which provides a new fusion protein for further experiments, the hJagged1(ext)-Fc fusion protein can be used as a new stimulator for proliferation of hematopoietic stem/progenitor cells in vitro.
Calcium-Binding Proteins ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Humans ; Immunoglobulin Fc Fragments ; biosynthesis ; genetics ; Immunoglobulin G ; biosynthesis ; genetics ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; genetics ; Jagged-1 Protein ; Membrane Proteins ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Serrate-Jagged Proteins ; Transfection

BACKGROUNDThe percutaneous transcatheter closure of secundum atrial septal defect (ASD) is increasingly a widespread alternative to surgical closure. The aim of this study was to assess long-term results of percutaneous closure of secundum-type atrial septal defect (ASDII).

METHODSBetween January 2001 and December 2005, 61 patients underwent a successful percutaneous closure of ASDII; including 25 male and 36 female. All were included in the patient study and were followed up to monitor by electrocardiogram and echocardiography, at intervals of 3 days, 3 months, 6 months, 1 year, 2 years, and 5 years after operation.

RESULTSThree days after percutaneous transcatheter septal closure (PTSC), the right atrium diameter, right ventricular end-diastolic left-right diameter and right ventricular end-diastolic volume (RVEDV) decreased significantly (P < 0.05). Right ventricular end-diastolic anteroposterior diameter (RVEDD), right ventricular end-systolic volume (RVESV) and right ventricular ejection fraction (RVEF) also decreased (P < 0.01). During the period from 3 to 6 months, the size of the right atrium and right ventricle returned to normal range. Three days after PTSC, the left ventricular end-diastolic diameter (LVEDD), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular-systolic volume (LVSV) and left ventricular ejection fraction (LVEF) were significantly increased (P < 0.05). At 1 year, the size of the left atrium, left ventricle and left cardiac function returned to normal range (P < 0.01). There were no deaths or significant complications during the study. At five year follow-up, all defects were completely closed and remained closed thereafter.

CONCLUSIONTranscatheter closure of ASDII effectively eliminated the abnormal shunt and, subsequently improved the dimensions of each chamber and cardiac function.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Heart Septal Defects, Atrial ; diagnostic imaging ; surgery ; Humans ; Male ; Middle Aged ; Ultrasonography ; Young Adult

Acupuncture Therapy ; Humans ; Medicine, Chinese Traditional ; Moxibustion ; Osteoporosis ; diagnosis ; pathology ; therapy ; Practice Guidelines as Topic

Objective To analyze the causes of missed diagnosis and misdiagnosis of abdominal organ le -sions through reviewing preoperative transabdominal ultrasound reports .Methods Data of the patients who re-ceived abdominal operation for abdominal organ lesions ( including liver , gallbladder , biliary tract , pancreas , spleen, kidney, adrenal gland, and appendix) in Peking Union Medical College Hospital within the period from March 1 to August 31 in 2013 were exported from pathological workstation .The preoperative ultrasound reports of these patients were reviewed .The missed diagnosis and misdiagnosis cases were recorded , and causes of the mis-takes were analyzed .Results Altogether 58 cases of missed diagnosis or misdiagnosis were identified from 1081 ultrasound reports (5.37%, 58/1081), including 6 liver lesions (5.77%, 6/104, all misdiagnosed), 6 gall-bladder and biliary tract lesions ( 1.30%, 6/462 , 5 missed and 1 misdiagnosed ) , 14 pancreatic lesions (19.72%, 14/71, all missed), 20 kidney and adrenal lesions (6.47%, 20/309, 11 missed and 9 misdiag-nosed), and 12 appendical lesions (16.00%, 12/75, 11 missed and 1 misdiagnosed).The average maximum diameter of the missed nodular lesions was significantly smaller than that of the misdiagnosed lesions ( P =0.001 ) .Conclusions Missed diagnosis and misdiagnosis of ultrasound are attributable to various causes , inclu-ding the nature , location , and size of abdominal organ lesions and the limitation of transabdominal ultrasound technology .The clinical ultrasound examination should be carried out very carefully and thoroughly .Ultrasound radiologists should have a thorough understanding of characteristics of different organ lesions and the limitation of ultrasound technique , in order to avoid missed diagnosis and misdiagnosis in clinical practice .

Eleven compounds were isolated from the 95% ethanol extract of the stems of Dendrobium officinale after water extraction by various modern chromatographic techniques, such as silica gel column chromatography(CC), octadecyl-silica(ODS) CC, Sephadex LH-20 CC, preparative thin layer chromatography(PTLC) and preparative high performance liquid chromatography(PHPLC). According to spectroscopic analyses(MS, 1D-NMR, 2D-NMR) combined with optical rotation data and calculated electronic circular dichroism(ECD), their structures were identified as dendrocandin Y(1), 4,4'-dihydroxybibenzyl(2), 3-hydroxy-4',5-dimethoxybibenzyl(3), 3,3'-dihydroxy-5-methoxybibenzyl(4), 3-hydroxy-3',4',5-trimethoxybibenzyl(5), crepidatin(6), alternariol(7), 4-hydroxy-3-methoxypropiophenone(8), 3-hydroxy-4,5-dimethoxypropiophenone(9), auriculatum A(10) and hyperalcohol(11). Among them, compound 1 was a new bibenzyl derivative; compounds 2 and 7-11 have not been previously reported from Dendrobium plants; compound 6 was reported from D.officinale for the first time. Compounds 3-6 exhibited potent antioxidant activity with IC_(50) values of 3.11-9.05 μmol·L~(-1) in ABTS radical scavenging assay. Compound 4 showed significant inhibitory effect on α-glucosidase, with IC_(50) value of 17.42 μmol·L~(-1), indicating that it boasted hypoglycemic activity.
Dendrobium ; Biological Assay ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Bibenzyls