OBJECTIVETo 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.

METHODSHuman 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).

RESULTSIn 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 significant differences 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.

CONCLUSIONSIn 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.

Alkaline Phosphatase ; metabolism ; Cell Culture Techniques ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Insulin-Like Growth Factor I ; pharmacology ; Osteocalcin ; genetics ; metabolism ; Osteogenesis ; drug effects ; Periodontal Ligament ; cytology ; RNA, Messenger ; metabolism ; Stem Cells ; cytology ; metabolism

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

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

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

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

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

To evaluate the efficacy and safety of Lianhua Qingwen capsule for influenza. All reports of the randomized controlled trials (RCTs) on Lianhua Qingwen capsule treating influenza were retrieved from database of CNKI, WANFANG DATA, VIP, PubMed, the Cochrane Library by February 2017. The studies were screened according to the inclusion and exclusion criteria, the data were extracted by 2 authors, the quality of the included RCTs was assessed, and meta-analysis was performed using Revman5.3 software. A total of 1 525 patients and 10 studies were included. The results of meta analysis showed that compared with oseltamivir, Lianhua Qingwen capsule was more effective in alleviating flu symptoms, including the time of headaches disappeared [SMD=-0.25,95% CI(-0.48, -0.01)], the time of sore throat disappeared [SMD=-0.53,95% CI（-0.72, -0.34）], the time of cough disappeared [SMD=-0.39,95%CI（-0.57, -0.21）], whole body aches disappeared [ SMD=-0.49, 95% CI (-0.78, -0.21)], the time of weak disappeared [SMD=-0.56,95%CI (-0.82, -0.29)], and the time of abatement of fever [SMD=-3.47,95%CI（-6.27, -0.67）]. Also, there were some statistical significant differences between the two groups except nasal congestion and the time of virus turning negative. Compared with Ribavirin, Lianhua Qingwen capsule was more effective in terms of the rate of temperature effect, [RR=1.53, 95% CI (1.24, 1.90)], the difference between the two groups was statistically significant. Compared with Ankahuangmin capsules, significant differences were found in terms of the he rate temperature effect [RR=1.37, 95%CI (1.19,1.57)]. Current evidence shows that Lianhua Qingwen capsule is more effective and safer than Oseltamivir, Ribavirin and Ankahuangmin capsules. Due to the low quality of the clinical research, the accuracy of this conclusion needs to be conducted to verify.

Objective To investigate the correlation between ultrasound-guided diffuse optical tomography (US-DOT) and hypoxia-inducible factor-1Α (HIF-1Α) of breast cancer. Methods Totally 69 patients with pathologically confirmed breast cancer underwent preoperative conventional breast ultrasonography examinations and US-DOT at Peking Union Medical College Hospital From October 2007 to February 2010 were enrolled in this study.After surgery,immunohistochemical staining of HIF-1Α and CD34 were performed,and the differences of total hemoglobin concentration (THC) and microvessel density (MVD) between HIF-1Α positive and negative groups were analyzed. Results HIF-1Α was positive in 12 cases (17.4%) and negative in 57 cases (82.6%). The average THC and MVD of HIF-1Α-positive cases were (274.763±77.661) Μmol/L and (33.8±10.8)/0.2 mm(2) respectively. The average THC and MVD of HIF-1Α-negative cases were (228.059±65.760)Μmol/L and (28.4±7.4)/0.2 mm(2). MVD(t=2.049,P=0.04) and THC(t=2.167,P=0.034) of HIF-1Α-positive group were significantly higher than those of HIF-1Α-negative group. Conclusions HIF-1Α can promote tumor angiogenesis and thus increase the blood supply and THC. As an indicator of tumor blood supply,THC can indirectly reflect the angiogenic activity of breast cancer.
Breast Neoplasms ; diagnostic imaging ; metabolism ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Neovascularization, Pathologic ; Tomography, Optical ; Ultrasonography, Mammary