OBJECTIVETo investigate the influence of the diameter and length of the mini-implant on the primary stability after loading with composite forces (CF) which contained torque and horizontal forces (HF).

METHODSNinety-six finite element models were established by the combination of mini-implant and bone, diameters (1.2 mm, 1.6 mm, 2.0 mm) and length (6 mm, 8 mm, 10 mm, 12 mm). There were 12 sizes, each size corresponded with 8 models. Group HF (each size n = 4) was loaded with 1.96 N horizontal force and Group CF (each size n = 4) was loaded with composite force which contained 6 N·mm torque and 1.96 N horizontal force. The maximum displacement of mini-implant with different force directions, implant diameters and lengths were evaluated.

RESULTSThe effect of force direction on the displacement related to diameter of mini-implant. The maximum displacement under load with HF respectively was changed with the changing of diameter[1.2 mm: (7.71 ± 0.49) µm; 1.6 mm: (3.94 ± 0.31) µm; 2.0 mm: (2.32 ± 0.43) µm], which were smaller than the maximum displacement of Group CF [1.2 mm: (9.22 ± 0.63) µm; 1.6 mm: (4.62 ± 0.52) µm; 2.0 mm: (2.69 ± 0.49) µm] (P < 0.05). When diameter was 1.2 mm, the difference of the maximum displacement [(1.61 ± 0.22) µm] between Group HF and CF was more obvious than that when the diameter was 1.6 mm or 2.0 mm [(0.64 ± 0.12), (0.49 ± 0.06) µm] (P < 0.05).

CONCLUSIONSThe composite force had unfavorable effect on the primary stability of the mini-implant. The diameter of the mini-implant had better be larger than 1.2 mm when the composite forces were applied.

Finite Element Analysis ; Orthodontic Anchorage Procedures ; instrumentation ; Torque

The study was aimed to evaluate if the modified in situ cryopreservation could affect the biological function of mesenchymal stem cells (MSC) in vitro. Mesenchymal stem cells from human bone marrow were isolated by standard method and characterized with their morphology, cell-surface antigen profile and differentiation repertoire in vitro. The culture-expanded MSC were cryopreserved in situ with culture medium (DMEM-LG) containing 10% D MSO and 30% selected FCS in -70 degrees C. Following recovery of cryopreservation, differentiation to adipocytes, chondrocytes, and osteoblast in vitro and cell cycle analysis were performed to investigate whether the cryopreservation would change the differentiation potential of MSC. The results showed that after recovery of cryopreservation, there was no changes detected as compared with the culture-expanded MSC in both differentiation potency and growth pattern at 12 weeks. In conclusions: this optimized short term in situ cryopreservation at -70 degrees C could retain biological characteristics of human MSC for at least 3 months, and this method may be useful for cryopreservation of hum an bone marrow MSCs.
Bone Marrow Cells ; cytology ; Cell Cycle ; Cell Differentiation ; Cell Separation ; Cell Survival ; Cryopreservation ; Humans ; Mesenchymal Stromal Cells ; cytology

OBJECTIVETo study the effect of angiogenesis inhibitor YH-16 in combination with 5-FU on liver metastasis of colorectal cancer.

METHODSIn vitro, the inhibitory effects of YH-16 and 5-FU on the growth of vascular endothelial cells and colorectal cancer cells were examined by MTT assay. In vivo, colorectal cancer cells were transplanted into BALB/c mice, and the mice were divided into six groups randomly:control group, low-dose YH-16 group, middle-dose YH-16 group, high-dose YH-16 group, 5-FU group and combination group. The number of liver metastases, the size of primary tumor and the toxicity were examined after 2 weeks postoperatively. The expression of vascular endothelial growth factor (VEGF) in liver metastases was detected by immunohistochemistry, and tumor microvessel density (MVD) was measured by immunostaining with CD34 and factor VIII (monoclonal antibodies.

RESULTSIn vitro, YH-16 inhibited the growth of colon cancer cells and vascular endothelial cells, with the IC50 at (2.16+/-0.28) microg/ml and (0.64+/-0.10) microg/ml respectively. In vivo high-dose YH-16 and 5-FU had a remarkable inhibitory effect on liver metastasis, and the combination group showed significant enhancement on this effect (P< 0.05). The combination group and 5-FU group could inhibit the growth of primary tumor, but not found in YH-16 group. The toxicity of YH-16 was lower than that of 5-FU (P< 0.05), and the difference was not found in the toxicity between combination group and 5-FU group (P > 0.05). Expression of VEGF in liver metastases was clearly inhibited by YH-16 in combination with 5-FU or 5-FU alone compared to the control group, and MVD in middle-dose and high-dose YH-16 group, 5-FU group and combination group was lower than that in control group (P< 0.05).

CONCLUSIONSThe angiogenesis inhibitor YH-16 can inhibit liver metastasis of colorectal cancer through inhibiting the growth of vascular endothelial cells. YH-16 in combination with 5-FU has additive effect on inhibitory activity against liver metastasis.

Angiogenesis Inhibitors ; therapeutic use ; Animals ; Cell Line, Tumor ; Colorectal Neoplasms ; drug therapy ; pathology ; Drug Therapy, Combination ; Female ; Fluorouracil ; therapeutic use ; Liver Neoplasms ; prevention & control ; secondary ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo construct an eukaryotic expression vector for PRL-2 and evaluate its effect on the invasiveness and migration of a human hepatocellular carcinoma cell line.

METHODSRT-PCR was performed to amplify the complete PRL-2 open reading frame using the total mRNA of hepatocellular carcinoma HepG2 cells as the template. PRL-2 gene was inserted into the pGEM T easy vector and sequenced, and the correct PRL-2 sequence was subcloned into the mammalian expression vector pcDNA3.1+. The constructed PRL-2 vector was transfected into CL1 cells via lipofectamine, and the stable expression of PRL-2 mRNA was detected by RT-PCR, the expressed protein identified by immunohistochemistry and Western blotting, and the effect of PRL-2 on the adhesion ability of CL-1 cell evaluated with MTT assay 20 and 120 min after transfection. The effect of PRL-2 on the invasive migration of CL-2 cells was evaluated according to the number of cells penetrating the Matrigel layer of polycarbonate membrane of Boyden chamber.

RESULTSRT-PCR yielded a fragment of 504 bp and the inserted PRL-2 sequence was verified by sequence analysis. The subclones were identified by restriction endonuclease digestion, and a G418-resistant clone, PRL-2-CL1, was obtained after 8 weeks of selection. RT-PCR showed stable expression of PRL-2 mRNA, and Western blotting confirmed overexpression of PRL-2 protein in the transfected cells. PRL-2 increased the adhesion rate of CL-1 cells to fibronectin at 20 min and 120 min after transfection (P<0.05), and also the number of CL-1 cells penetrating the polycarbonate membrane from 10.0+/-3.7 to 44.8+/-2.6 (P<0.05).

CONCLUSIONAn eukaryotic expression vector of PRL-2 has been successfully constructed, which allows stable and efficient expression in CL-1 cell line. PRL-2 can promote cell adhesion and invasion activity of human hepatocellular carcinoma cells.

Animals ; Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Adhesion ; genetics ; Cell Line, Tumor ; Cell Movement ; genetics ; Eukaryotic Cells ; metabolism ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Humans ; Liver Neoplasms ; genetics ; pathology ; Neoplasm Invasiveness ; genetics ; Protein Tyrosine Phosphatases ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo study the effect of skin-derived progenitor cell (SKP) combined with hyaluronic acid( HA) on the wound healing in diabetic rats.

METHODSSKP of Spraque-Dawley (SD) neonate rats were isolated and cultured and mixed with HA. The differentiation characteristics of SKP in the culture were observed. Sixty SD rats were injected intraperitoneally with 65 mg/kg streptozotocin( STZ) to induce diabetes. Two symmetrical full-thickness cutaneous wounds( 1.0 cm in diameter) were made on the back of each SD rat and randomly divided into A (n = 20, with treatment of 100 mircol SKP-HA) , B (n = 20, with treatment of 100 mirol HA) , and C ( n = 20, with treatment of DMEM/F12 culture medium) groups. Tissue samples from wound in each group were harvested on 1, 2, 3, 4 weeks after the treatment. Wound healing rate, changes in histomorphology, the content of hydroxyproline ( HYP) , and immigration of labelled SKP were determined and analyzed.

RESULTSSKP grew well when cultured with HA. The characteristics of SKP to differentiate into lipocyte, neuron, and neurogliocyte remained in the culture. Compared with that in C group, epithelization in the wounds of A and B groups appeared earlier. The wound healing rate in A group [ (72.1 +/- 2. 8)% ] and B group [ (53.7 +/- 2. 9)% ] were obviously higher at 2 post-treatment weeks(PTW) than that in group C [(42. 5 +/- 1.5)% ( P <0.05) , and that in A group was obviously higher compared with B and C groups at 3 PTW ( P < 0. 05 or 0. 01). The wound healing rates in A and B groups were (100. 00 +/- 0.00) % at 4 PIW, which were obviously higher than that of group C( P <0.01) . There was no obvious difference in the HYP content among the 3 groups at 1 PIW ( P > 0. 05) , but it was obviously higher in A and B groups than that in C group at 2,3,4 PTW( P <0.01) , and that in A group was significantly higher than that in B group at 3 and 4 PTW( P <0. 01). SKP survived well on the wound, and migrated towards the dermis as time elapses.

CONCLUSIONSKP-HA composition can promote wound healing in diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; pathology ; Hyaluronic Acid ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Skin ; cytology ; Stem Cells ; chemistry ; cytology ; Wound Healing ; drug effects

OBJECTIVETo study effects of different intraabdominal pressure of carbon dioxide (Cq2) pneumoperitoneum on hemorrheology and microcirculation in rabbits.

METHODSEighteen female healthy rabbits weighing 2.2 kg to 3.5 kg were randomly divided into three groups equally based on pneumoperitoneum pressure: 0 mmHg group (group I),10 mmHg group (group II) and 15 mmHg (group III). Each group received 1 h pneumoperitoneum under different pressure. Blood samples were taken at 5 min before CO2 pneumoperitoneum, at 30 and 60 min after pneumoperitoneum for the measurements of indexes of hemorrheology. Hemodynamics including heart rate (HR), mean arterial pressure (MAP) and the volume and velocity of the microcirculation of auricle were continuously monitored, such indexes were recorded at the related time.

RESULTSAfer pneumoperitoneum at 30 and 60 min, compared with group I, HR, MAP, the whole blood viscosity, the aggregation and rigid indexes of RBC were significantly raised in group II (P < 0.05), the deformability indexes of RBC, the volume and velocity of the microcirculation were markedly decreased (P < 0.05). Even more significant changes were observed in group III (P < 0.01). The plasma viscosity and the hematocrit changed little.

CONCLUSIONAfter CO2 pneumoperitoneum, hemorrheology is decreased; Although HR, MAP are raised, the volume and velocity of the microcirculation are decreased.

Abdomen ; blood supply ; Animals ; Blood Viscosity ; Carbon Dioxide ; Female ; Hematocrit ; Hemorheology ; Microcirculation ; Pneumoperitoneum, Artificial ; methods ; Pressure ; Rabbits

OBJECTIVETo evaluate the learning process of video-assisted minimally invasive esophagectomy (MIE).

METHODSOne hundred consecutive patients with thoracic esophageal carcinoma were treated by a same team of surgeons, and were divided into 3 groups in chronological order. The former two groups both consisted of 25 patients with thoracoscopy plus laparotomy. The remaining 50 patients were enrolled in the third group with thoracoscopy plus laparoscopy. Clinicopathological data including operative time, blood loss, protection of normal structures, complications, length of ICU stay, postoperative stay, and lymph nodes harvest, were collected and compared between groups.

RESULTSProcedures were accomplished successfully in 96 patients. Only 4 cases were converted to open thoracotomy and none to laparotomy. The median operative time was 310 min and blood loss was 200 ml. The median number of lymph node harvest was 22. The overall complication rate was 50%. Comparison of first two groups revealed that significant differences existed in the preservation rate of arch of azygos vein (P=0.010), bronchial vessels (P=0.038), and exposure rate of thoracic part of left recurrent laryngeal nerve( P=0.048). Comparison of the former and latter 50 patients revealed that significant differences existed in thoracic operative time (P<0.001), blood loss (P=0.025), preservation rate of arch of azygos vein (P=0.001) and bronchial vessels (P<0.001), the number of lymph node harvest in thoracoscopy (P=0.022) and in left recurrent laryngeal nerve chain (P<0.001), and exposure rate of initiate part of left recurrent laryngeal nerve (P=0.002).

CONCLUSIONThe learning curve of MIE is long and beginners should proceed step by step.

Adult ; Aged ; Esophageal Neoplasms ; surgery ; Esophagectomy ; methods ; Female ; Humans ; Learning Curve ; Male ; Middle Aged ; Retrospective Studies ; Thoracoscopy ; methods ; Video-Assisted Surgery ; methods

OBJECTIVETo analyze the gene expression level of fibroblast activation protein in HBV related hepatocellular carcinoma patients and discuss its clinical significance.

METHODSFAP gene expression in 33 hepatocellular carcinoma patients cancer tissues, peficancerous tissues, distant relative normal liver tissues and 13 normal liver tissues were examined by reverse transcription PCR; and real-time fluorescent quantitative PCR (qRT-PCR) was used to quantify their expression.

RESULTSFAP were expressed in all the tissues,the relative expression values in cancer tissues, peficancerous tissues and distant relative normal liver tissues were 5.14 +/- 6.69, 1.58 +/- 0.96, 1.63 +/- 0.94, respectively, the differences were statistically significant (F = 4.401, P < 0.05); and in TNM stage I, II, IIII, they were 2.89 +/- 3.35, 4.15 +/- 4.69, 10.09 +/- 9.51 respectively; in well-differentiated, differentiated and poorly differentiated hepatocellular carcinoma were 1.62 +/- 1.74, 3.84 +/- 3.79, 1.26 +/- 13.34 respectively. The differences were all statistically significant (P < 0.05).

CONCLUSIONFAP may play an important role in the occurrence and development of HBV related hepatocellular carcinoma.

Adult ; Aged ; Carcinoma, Hepatocellular ; etiology ; metabolism ; Female ; Gelatinases ; genetics ; physiology ; Gene Expression Regulation, Neoplastic ; Hepatitis B ; complications ; Humans ; Liver Neoplasms ; etiology ; metabolism ; Male ; Membrane Proteins ; genetics ; physiology ; Middle Aged ; RNA, Messenger ; analysis ; Serine Endopeptidases ; genetics ; physiology

OBJECTIVETo summarize the management of anastomotic leak following surgery for esophageal carcinoma.

METHODSThe medical records of the patients developing digestive tract leak after surgery for esophageal carcinoma in our hospital from January 2003 to March 2011 were retrospectively analyzed.

RESULTSA total of 36 patients were included, in whom 13 developed cervical anastomotic leak, 18 had intra-thoracic anastomotic leak, and 5 had intra-thoracic gastric necrosis. Of these patients, 7 were treated with resurgery, 6 with esophageal stent implantation, and 23 with conservative treatment. Treatment lasted for 5 to 181 days, averagely 47.0 +/- 31.9 days. After management, 9 patients died (25.0%). Among seven patients with resurgery, four had deceased, two were cured, and one developed leak again and was switched to conservative treatment until discharged. All the 6 patients treated with stent implantation were cured. Of the 24 patients receiving conservative treatment (including one switched from resurgery), 18 (75.0%) were cured and 1 was not cured but survived.

CONCLUSIONSAnastomotic leak following surgery for esophageal carcinoma should be treated individually based on the onset time, location, size, and extent of the leakage. Conservative treatment is still a safe and effective method. The efficacy of stent implantation needs further investigation to confirm.

Adult ; Aged ; Anastomotic Leak ; therapy ; Esophageal Neoplasms ; surgery ; Humans ; Male ; Middle Aged ; Precision Medicine ; Treatment Outcome

OBJECTIVETo investigate the protective effect of Astragalus Membranaceus Injection on human umbilical vein endothelial cells (HUVECs) against tumor necrosis factor alpha (TNF-alpha).

METHODSCultured passage 2 HUVECs were stimulated with TNF-alpha with or without a 2-h Astragalus Membranaceus Injection treatment. The expression of nuclear factor-kappaB (NF-kappaB) subunit p65 were evaluated by immuncytochemistical method, and the levels of p65 in the nuclei and the protein Ikappabetaalpha in the cytoplasm were evaluated by Western blotting. The levels of interleukin-6 (IL-6) and soluble intracellular adhesion molecule-1 (sICAM-1) in the cell culture were determined with ELISA.

RESULTSTNF-alpha induced the activation of NF-kappaB and increased the expressions of IL-6 and sICAM-1 in HUVECs. The activation of NF-kappabeta by TNF-alpha was suppressed by Astragalus Membranaceus Injection in a dose-dependent manner.

CONCLUSIONAstragalus Membranaceus Injection can inhibit the TNF-alpha-induced expression of IL-6 and sICAM-1 by suppressing NF-kappabeta activation, suggesting its protective effect on the endothelial function.

Astragalus membranaceus ; chemistry ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; cytology ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-6 ; metabolism ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; pharmacology ; Umbilical Veins ; cytology