OBJECTIVE: To investigate the biological characteristics of endothelial progenitor cells (EPCs) from the umbilical cord blood (UCB), and to evaluate their oncogenicity after long-term culture in vitro. METHODS: The mononuclear cells (MNCs) were isolated from the UCB and cultured in MCDB131 medium supplemented with 20% FBS, VEGF and other growth factors. Morphology of the EPCs was observed, and the growth curve of the EPCs was investigated. Surface antigens of the EPCs were analyzed by the flow-cytometer. The capability of intaking the acetylated low-density lipoprotein (acLDL) of the EPCs was detected using fluoresencent chemical method. The vasoformative capability and genetic stability of EPCs were cultured in matrigel, and examined by karyotype analysis. The oncogenicity of EPCs was verified by the tumorigenesis test in athymic mouse and soft agar. RESULTS: EPCs were successfully derived from the UCB, and could be passaged to at least 42(nd) generation and had strong abilities of proliferation, acLDL intake and vasoformation, but there was not oncogenicity. They expressed endothelial cell-surface antigens and maintained normal karyotype. CONCLUSION The EPCs with proliferative potential can be isolated from the UCB. They can be passaged in long-term cultures without oncogenicity, and can maintain normal karyotype. The EPCs can be served as a new type of cells in cell and gene therapy.
Animals ; Antigens, Surface ; analysis ; Cell Line ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Fetal Blood ; cytology ; Flow Cytometry ; HeLa Cells ; Humans ; Infant, Newborn ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Karyotyping ; Mice ; Mice, Nude ; Neoplasms, Experimental ; pathology ; Stem Cells ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo investigate the mechanism of new bone formation in the distraction osteogenesis (DO) for correction of cleft palate (CP) in rhesus.

METHODSCP was created by operation in 23 rhesus. The CP was corrected with DO in 21 animals as experimental group. The distraction rate was 0.8 mm per day, two times a day. The bone fragments were fixed after cleft closure, every 3 animals were sacrificed to get specimen after 1, 2, 4, 6, 8, 12, 24 weeks of fixation. 6 days before sacrifice, tetracycline was administrated for labeling (30 mg/kg).

RESULTSThe hard and soft tissue def of fixation. At the same time, the bone volume and calcification between the distraction gap increased. The cleft in the control group could not b ect was successfully closed with DO by intramembrane osteogenesis. The new formed bone was remodeling and became maturation during the period e corrected spontaneously.

CONCLUSIONSThe DO can successfully correct both the soft and hard tissue defect in CP by intramembrane osteogenesis. The fixation is important for remodeling and maturation of the new formed bone.

Animals ; Biomarkers ; Cleft Palate ; pathology ; surgery ; Macaca ; Osteogenesis, Distraction ; Palate, Hard ; pathology ; Palate, Soft ; pathology

OBJECTIVETo study the mechanism of new bone formation and remodeling of distraction osteogenesis (DO) by analysis of the expression of osteopontin (OPN) and osteocalcin (OC).

METHODSRhesus were operated to reconstruct the animal model of cleft palate (CP). The CP was closed by DO in experimental group(n = 21). After consolidation of 1, 2, 4, 6, 8, 12, 24 weeks, every 3 animals were killed to collect the specimens, respectively. The OPN and OC and their mRNA were detected quantitatively by Real-time RT-PCR and ELISA, respectively. The animals in control group (n = 2) and sham group (n = 2) were used as control.

RESULTSThe mRNA expression of OPN increased since 2nd week of consolidation and reached the peak at 4th week (7.59 +/- 0.37). The mRNA expression of OC was up-regulated since 4th week, and reach the peak at 6th week (7.94 +/- 0.31). Then they decreased to about the level in sham group at 24th week (P > 0.05). The OPN and OC were highly expressed during 4 to 6 weeks of consolidation. During 8 to 12 weeks, they decreased like their mRNA expression.

CONCLUSIONThe intramembraneous new bone formation after DO can reconstruct the bone defect of CP. The new formed bone can be remodeled to be quite normal bone tissue.

Animals ; Cleft Palate ; metabolism ; surgery ; Macaca mulatta ; Osteocalcin ; metabolism ; Osteogenesis, Distraction ; Osteopontin ; metabolism

OBJECTIVETo investigate the effects of Bushen Huoxue Fang (BSHX) on the apoptosis of epithelial cells in the prostatic ductal system of rats with benign prostatic hyperplasia (BPH) and its possible action mechanism.

METHODSOne hundred 3- month-old male Wistar rats were randomly divided into four groups of equal number (control, castrated, BPH model, and BSHX). BPH models were made by subcutaneous injection of testosterone following castration; the rats in the BSHX group were treated intragastrically with BSHX at 2.34 g/ml after modeling, while those in the other two groups with equal volume of saline, all for 37 days. On the 38th day, all the rats were sacrificed and their prostates harvested for detection of the distribution of TGF-beta1 and alpha-actin and the count of positive cells in the prostatic ductal system by immunohistochemical staining. The apoptosis rate of epithelial cells in the prostatic ductal system was determined by TUNEL assay.

RESULTSThe expression of TGF-beta1 was significantly increased in the rats of the BSHX group as compared with the BPH models in both the proximal prostatic duct ([15.28 +/- 4.30]% vs [36.42 +/- 8.10]%, P < 0.01) and the distal prostatic duct ([4.42 +/- 2.07]% vs [8.71 +/- 2.28 ]%, P < 0.05), while the expression of alpha-actin in the proximal duct was remarkably higher in the BSHX-treated rats than in the models ([28.14 +/- 7.43]% vs [18.28 +/- 4.07]%, P < 0.01), but lower than in the control animals ([33.57 +/- 6.85]%, P < 0.05). Compared with the control group, the BPH models and BSHX-treated rats both exhibited markedly decreased apoptosis of epithelial cells in the proximal prostatic duct ([39.42 +/- 9.20]% vs [3.86 +/- 1.34]%, P < 0.01, and [31.14 +/- 5.64]%, P < 0.01) and distal prostatic duct ([17.60 +/- 4.86]% vs [3.07 +/- 1.14]%, P < 0.01, and [12.37 +/- 2.25]%, P < 0.05). The apoptosis rate of epithelial cells in the prostatic ductal system was significantly higher in the BSHX-treated rats than in the BPH models (P < 0.01).

CONCLUSIONBy upregulating the expression of TGF-beta, BSHX can suppress the reduction of smooth muscle cells in the proximal prostatic duct, promote the apoptosis of prostatic epithelial cells, and thus effectively inhibit benign prostatic hyperplasia.

Actins ; metabolism ; Animals ; Apoptosis ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; pathology ; Male ; Prostatic Hyperplasia ; drug therapy ; metabolism ; pathology ; Rats ; Rats, Wistar ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo study the ultrastructure and Ca/P element spectrometry of distraction osteogenesis (DO) for reconstruction of cleft palate (CP), so as to explore the osteogenesis and remodeling of new bone in situ.

METHODS23 rhesus macaques were operated to establish animal models of CP. 2 monkeys didn't received DO as controls. The other 21 monkeys in experimental group underwent DO to correct both bony and soft tissue defects in palate. The distraction was performed at a rate of 0.8 mm/d, twice a day until the cleft was closed. After fixation for 1, 2, 4, 6, 8, 12, 24 weeks, every 3 animals were sacrificed to get the specimens at the distraction gap. The scanning electron microscopic study and Ca, P elements spectrometric analysis were adopted. There were also two unoperated animals as sham group.

RESULTSAfter fixation for 1-2 weeks, the distraction gap was full of collagen fibers oriented along vector of distraction. Few trabeculae was seen at the margin area. After fixation for 4-6 weeks, active osteogenesis was presented with new formed bone trabeculae and abundant cellular component. After fixation for 8-12 weeks, the new formed bone became mature and couldn't distinguish from the normal bone. 24 weeks later, the bone between the distraction gap had a similar structure to the normal bone. Elements spectrometric analysis results indicated that in early stage of osteogenesis, the P and S peaks were relatively high while the Ca peak was much lower. During the late stage, the S peak was obviously decreased, and Ca/P ratio increased to normal level as in the empty control group.

CONCLUSIONSThe CP can be corrected by DO. The new bone between the distraction gap is formed and remodeled through intramembraneous osteogenesis.

Animals ; Cleft Palate ; metabolism ; pathology ; surgery ; Disease Models, Animal ; Female ; Macaca mulatta ; Male ; Microscopy, Electron, Scanning ; Osteogenesis ; Osteogenesis, Distraction ; methods ; Palate ; surgery ; ultrastructure

OBJECTIVETo study the value of multi-slice spiral computed tomography (CT) in the preoperative evaluation of lymph node metastasis.

METHODSTotally 45 patients with gastric cancer detected by 64-slice spiral CT were enrolled in this study. The potential lymph node metastasis was evaluated by measuring or calculating the long diameter, extent of enhancement, and short-to-long diameter ratio of the lymph nodes. The results were compared with postoperative pathological findings.

RESULTA long diameter ≥ 8mm,enhanced density ≥ 80Hu, and short-to-long diameter ratio ≥ 0.7 had the best consistency with postoperative pathological findings.

CONCLUSIONAs a simple and noninvasive technique, multi-slice spiral CT is helpful to identify potential lymph node metastasis of gastric cancer based on long diameter, extent of enhancement, and short-to-long diameter ratio of the lymph nodes, and thus provide important information for surgery selection, prognosis, and development of new procedures.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymph Nodes ; diagnostic imaging ; pathology ; Lymphatic Metastasis ; diagnostic imaging ; Male ; Middle Aged ; Stomach Neoplasms ; diagnostic imaging ; pathology ; Tomography, Spiral Computed

OBJECTIVETo observe the protective effect of Shenfu Injection (SFI) pretreatment on brain of patients receiving aortic valve replacement (AVR) undergoing cardiopulmonary bypass (CPB).

METHODSThirty AVR patients undergoing CPB were randomly assigned to 2 groups, the control group and the experimental group, 15 cases in each group. SFI at 1.5 mL/kg (dissolved in 250 mL 5% glucose solution) was intravenously dripped to those in the experimental group 5 days before operation, once daily for 5 successive days. SFI at 1.5 mL/kg (dissolved in 250 mL 5% glucose solution) was intravenously dripped to those 30 min before anesthesia induction. Equal dose of normal saline was intravenously dripped to those in the control group, and the other procedures were the same as those for patients in the experimental group. The venous blood sample (2 mL) was drawn from the right internal carotid vein immediately after induction of anesthesia (T1),10 min after CPB (T2), 30 min after GPB (T3), 2 h after CPB (T4), 24 h after CPB (T5), and 48 h after CPB (T6), thus detecting the plasma levels of S100beta and neuron specific enolase (NSE). And patients' cognitive function was assessed with mini-mental state examination (MMSE) scale on the day before operation, the 2nd and the 7th day after operation.

RESULTSThere was no statistical difference in the levels of S1001 and NSE between the two groups at T1 (P > 0.05). There was statistical difference in the levels of S100beta and NSE between the two groups at T2, T3, T4, T5, T6, when compared with those at T1 (P <0.05). Besides, the levels of S100beta and NSE at T2, T3, T4, T5, T6 were lower in the experimental group than in the control group, showing statistical difference (P <0.05). The MMSE scores decreased on the 2nd day after operation in the two groups, showing statistical difference when compared with those on the day before operation (P <0.05). It was lowered more obviously in the control group. There was no statistical difference in the MMSE score between the 7th day post-operation and the day before operation (P >0.05).

CONCLUSIONSFI pretreatment had protective effect on brain in AVR patients undergoing CPB.

Brain ; drug effects ; metabolism ; Cardiopulmonary Bypass ; Cognition ; drug effects ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Heart Valve Prosthesis Implantation ; methods ; Humans ; Intraoperative Period ; Ischemic Preconditioning ; methods ; Male ; Middle Aged ; Phosphopyruvate Hydratase ; metabolism ; S100 Calcium Binding Protein beta Subunit ; metabolism

Objective To investigate health-related quality of life (HRQOL) and influencing factors in adult patients with Kashin-Beck disease (KBD) in Shaanxi Province for improving health-related quality of life of KBD patients.Methods By the two-stage cluster random sampling method,data of 901 cases of KBD patients with the disease severity of grade 1 and higher were collected from higher KBD incidence areas of Baoji,Xianyang and Tongchuan in Shaanxi Province in February 2016.The Chinese version of SF-36 (including 8 dimensions:physical functioning,role-physical,bodily pain,general health,vitality,social functioning,role-emotional,mental health) was used to measure the HRQOL.Multiple linear regression was used to analyze the influencing factors of HRQOL.Results In 901 cases of KBD patients,male accounted for 55.49％ (500/901),female accounted for 44.51 (401/901).The median age of patients was 58 (22-83) years old.According to the severity of disease criteria,patients with grade Ⅰ were 37.18％ (335/901),patients with grade Ⅱ were 53.50％ (482/901) and patients with grade Ⅲ] were 9.32％ (84/901).The scores of each dimension of SF-36 scale in patients of male patients had higher scores in GH and MH than women [GH:35 (25,50) vs 30 (20,45),MH:56 (48,72) vs 56 (44,68),.9 ＜ 0.05].There were significant differences in PF,RP,BP,GH,VT and SF scores among different age groups (P ＜ 0.05).Similarly,there were significant differences in PF,RP,GH,VT,SF,RE and MH scores among patients with different educational levels (P ＜ 0.05).Married group was only higher in the PF and SF scores than other marital status groups [PF:55 (40,75) vs 50 (35,65),SF:62 (50,75) vs 50 (38,75),P ＜ 0.05].As expected,there were significant differences in the scores of PF,BP,GH and SF between the severity of diseases.There were also significant differences in the scores of PF,BP,GH,VT,SF and MH in KBD patients from different regions (P ＜ 0.05).According to multiple linear regression analysis results,male was the conservation factor of GH and MH scores compared with female;age was a risk factor for PF,RP,BP,GH,VT and SF scores;compared with the illiterate,primary school education was a protective factor for PF and RE scores,junior high school and above were PF,RP,VT,SF,RE and MH scores protective factors;compared with degree Ⅰ KBD,degree Ⅱ and degree Ⅲ were risk factors for PF,BP,GH and SF scores;compared with Baoji area,Xianyang area was a protective factor of SF score,however,Tongchuan area was a protective factor for PF,GH,VT,SF and MH scores.Conclusions KBD has severely damaged the HRQOL of patients.Priority should be given to age,educational attainment and severity of the disease for developing measures to improve KBD patients' HRQOL in different regions.

Objective To investigate the level of perceived social support and its influencing factors in Kashin-Beck disease (KBD) patients in Shaanxi Province,and to provide a theoretical basis for improving the service to KBD patients.Methods By the two-stage cluster random sampling method,KBD patients with the severity of disease of grade Ⅰ and above were investigated in Baoji,Xianyang and Tongchuan areas in Shaanxi Province,which were higher incidence areas of KBD.The Chinese version of perceived social support scale (PSSS) was used to measure family support,friend support,and other supportive comprehension social support levels by household survey method,the total score of PSSS is low support state from 12 to 36,intermediate support state from 37 to 60,and high support state from 61 to 84,and the effects of gender,age,education level,marital status,occupation,disease severity and region on the level of perceived social support were analyzed.Results In 901 cases of KBD patients,males accounted for 55.49％ (500/901),females accounted for 44.51％ (401/901).The median age of patients was 58 years old.According to the severity of KBD criteria,grade Ⅰ patients were 37.18％ (335/901),grade Ⅱ patients were 53.50％ (482/901) and grade Ⅲ patients were 9.32％ (84/901).The median (quartile) PSSS score was 62 (54,70),the low support status accounted for 8.55％ (77/901),the intermediate support status accounted for 36.96％ (333/901),and the high support status was 54.50％ (491/901).Age was a risk factor for friend support,other support dimensions and total score;compared with illiteracy,primary school education was the protective factor of total score,and junior high school and above education was the protective factor of family support dimension and total score.Compared with Baoji area,Xianyang area and Tongchuan area were the protective factors of family support dimension,other support dimension,and total score.In addition,Xianyang area was also a protective factor of friend support dimension.Conclusion The level of perceived social support of KBD patients in Shaanxi is generally above moderate.Age,education,and regional factors are main factors affecting perceived social support.

Objective To investigate the neuroprotective effect of ischcmic posteonditioning (IP)against cerebral ischemia-reperfusion injury and the role of phosphoinositide 3-kinase(P13K)signaling pathway in the neuroprotection. Methods Focal cerebral ischernia was induced in 24 SD rats by permanent distal middle cerebral artery occlusion and transient bilateral comlllOn carotid artery occlusion.The rats were then randomized into 4 groups for treatment with IP,LY294002+IP,DMSO+IP,or without IP.In LY294002+IP and DMSO+IP groups,LY294002 or DMSO was injcoted into the ventricular space on the ischemic side 1 h before ischemia.The cerebral infarct sizes wgre measured in all the 4 groups at 48h after the reperfusion.Results Cerebral infarcts were observed in all the groups on theischemic side,all locating in the left neocortex and the middle cerebral artery territory.At48h after reperfusion,the infarct size was significantly smaller in rats with IP(34.02%±7.17%)than in those without IP(57.05%±10.05%)(P＜0.05),and significantly larger in LY294002+IP group(73.41%±2.06%)than in DMSO+IP group(35.76%±1.51%)(P＜0.05).No significant difference was found in the infarctsize between DMSO+IP group and IP group(P＞0.05).Conclusion IP ameliorates cerebral reperfusion mjury in rats,and the mechanism of this neuroprotective effect involves the preservation of PI3K activity.