Objective:To isolate and identify the periodontal ligament stem cells.Methods:Periodontal ligament tissues were digested with a mixture of collagenase and Dispase,then the periodontal ligament stem cell clones were isolated by limited dilution method.The transmission electron microscopy(TEM),flow cytometery and immunohistochemistry procedure were employed to study the ultrastructure,cell cycle and surface marker of the cloned cells.Results:The obtained cells showed the characteristics of undifferentiated morphology at ultrastructural level.94.4% of the cells were in phase G_0/G_1.The cells were Vimentin,STRO-1,ON and BSP positive.Conclusion:Cloning incubation may be the effective way to isolate and purify the periodontal ligament stem cells.

OBJECTIVE:To simultaneously determine the contents of CIPRO and MNZ in compound ciprofloxacin ear dro_ ps by nodical polyploid UV-spectrophotometry.METHODS:CIPRO and MNZ were nodical absorption at wavelength of350.0nm and this was just the sum of isoabsorptive point of two components while DXM did not absorb UV ray at this point.Therefore350.0nm was adopted as the detecting wavelength for these two components.RESULTS:The average recovery and relative standard deviation of CIPRO were99.7%and1.4%respectively.The average recovery and relative standard de?viation of MNZ were100.7%and1.6%respectively.CONCLUSION:The method is accurate,rapid,simple and sensitive.This method can be used to control the quality of this preparation.

Objective To analyze tissue vascular endothelial growth factor receptor 1 (VEGFR-1)and VEGFR-2 and their soluble form (sVEFGR-1 and sVEGFR-2) in the plasma in patients with preeclampsia,and to explore its clinical significance.Methods sVEGFR-1 and sVEGFR-2 expressions in plasma of normal pregnancy women and preeclampsia patients were detected by enzyme-linked immunosorbnent assay (ELISA) ; VEGFR-1,VEGFR-2,sVEGFR-1 and sVEGFR-2 mRNA expressions of normal pregnancy women and preeclampsia patients in placenta were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results ELISA results showed that the level of sVEGFR-1 in plasma of normal control group before and after delivery was (12.33 ± 1.52) ng/ml and (4.55 ± 0.31) ng/ml,respectively,while that of preeclampsia group was (77.25 ± 9.47) ng/ml and (8.13 ± 0.74) ng/ml,respectively; the differences between before and after delivery in the two groups were of statistical significance.The level of sVEGFR 2 in plasma of normal control group before and after delivery was (8.74 ± 1.24) ng/ml and (6.43± 0.55) ng/ml,respectively,while that of preeclampsia group was (5.69 ± 0.75) ng/ml and (4.96 ±0.67) ng/ml,respectively.RT-PCR results were consistent with ELISA results.Conclusions Rapid decrease of plasma sVEGFR-1 and continuously low-level expression of plasma sVEGFR-2 indicated that VEGFR-1 might be closely related to preeclampsia,and decrease of plasma sVEGFR-2 in preeclampsia women might be taken as a marker of endothelial cell function disorder.

Objective To evaluate the clinical application and effect of plastic surgery in emergency treatment of facial soft tissue injuries,and to explore the better plastic surgery method for facial soft tissue injuries in order to regain the patient facial morphology and function maximally.Methods The clinical data of 798 patients with facial soft tissue injuries from June 2009 to June 2011 were analyzed retrospectively.And plastic surgical techniques were applied to the early treatment of facial soft tissue injuries in this group cases,according to the size of defect and the degree of deformity of the patient,different plastic surgery treatment was chosen,such as skin flap or skin graft to repair wound surfaces.In this process,one must follow sterile noninvasive principle strictly with emphasis on the technique of plastic surgery such as entire debridement,wound healing application of skin flap and so on.Results 790 cases of facial soft tissue injuries were healed by first intention without significant complications,while 8 cases of them had mild scars.During 6 to 12 months of follow-up,neither scar,nor infections and necrosis of the wound region occurred,and the morphology and function of patients' face recovered well without the second operation.Conclusions Using plastic surgery techniques in the emergency treatment of facial soft tissue injuries as soon as possible can avoid the disfigurement and the function disturbance,and promote the facial morphology and function regeneration rapidly and effectively.

Objective To observe expressions of mRNA for Par-4 and WT1 in bone marrow cells from acute leukemia patients and non-leukemia patients, and to approach the correlation between CR rate and Par-4, WT1 expression level. Methods To detect Par-4 and WT1 mRNA expression level in bone marrow cells from 78 acute leukemia patients and 23 non-leukemia patients by means of Real-time Fluorescent Quantitation RT-PCR. Results FQ-RT-PCR result showed that Par-4 mRNA was expressed in bone marrow cells from 78 acute leukemia patients and 23 non-leukemia patients. Compared with control groups, the expression levels of Par-4 mRNA were significantly suppressed (9.35×10-4±8.4×10-5, P ＜0.05). Compared with initial treatment groups and relapse groups, the expression levels of Par-4 mRNA in remission groups were significantly up-regulated (1.26×10-3±1.1×10-4) but were still significantly lower than that in control groups (3.25×10-3±2.9×10-4). There was no significance difference between initial treatment groups and relapse groups. No apparent association was found between Par-4 expression level and CR rate (P ＞0.05). WT1 gene was overexpressed in bone marrow cells from acute leukemia patients(2.98× 10-3±2.1×10-4), but the expression levels of WT1 mRNA were significantly lower in bone marrow cells from control groups (7.25×10-5±6.7×10-6,P ＜0.05). Compared with initial treatment groups and relapse groups, the expression levels of WT1 mRNA in remission groups were significantly down-regulated (6.86×10-4±5.2× 10-5) but were still significantly higher than that in control groups. There was no significant difference between initial treatment groups and relapse groups.There was significant difference between different WT1 expression levels and CR rates (P ＜0.05). Conclusion The result of FQ-RT-PCR testing confirmed that Par-4 mRNA expression is lower, while WT1 is higher in acute leukemia. Par-4 and WT1 gene present mutually exclusive expression patterns. There was no apparent association between Par-4 expression level and CR rate.

Objective To explore the safety and efficacy of small margin in nephron sparing surgery for early localized renal cell carcinoma (RCC). Methods A total of 325 cases of RCC with normal contralateral kidney and staged as Tla were retrospectivly studied.According to the margin size,125 cases were with surgical margin ≤ 5 mm (group ≤ 5 mm),102 cases with margin 6-9 mm (group 6-9 mm) and 98 cases with margin ＞ 10 mm (group ＞ 10 mm).The margin size and status was pathologically evaluated and clinical results including local recurrence,distant metastasis and overall survival rate were followed up and comparatively analyzed. Results None of the patients had positive surgical margins.The mean and median margin sizes were 2.2 and 2.0 mm for group ≤ 5 mm,6.7 and 6.0 mm for group 6-9 mm and 11.8 and 12.0 mm for group ＞ 10 mm.The difference was statistically significant (P=0.025).The mean and median follow-up time for all the patients were 79 and 83 months (range 15-132 months),with 69 and 73 months (range,15-130 months) for group ≤ 5 mm,83 and 86 (range,17-132 months) for group 6-9mm and 82 and 82 (range 60-103 months) for group ＞ 10 mm.Three patients in group ≤ 5 mm,5 in group 6-9 mm and 2 in group ＞ 10 mm died of no-cancer related disease during follow-up.One patient in group ≤ 5 mm (0.74％) experienced ectopic recurrence in the same kidney and one in group 6-9 mm was detected local recurrence in situ (0.98％).No distant metastasis was detected in all the patients.The overall 5-year survival rate for patients in groups ≤ 5mm,6-9 mm and ＞ 10 mm were 99.2％,99.0％ and 98.0％,respectively.(Kaplan-Meier survival analysis,Log Rank,x2 =1.511,P=0.470). Conclusions Small margin in nephron sparing surgery is safe and effective in treating RCC with stage T1a,which provides excellent renal function preservation,favorable long-term progression-free survival rate,and is not associated with an increased risk of local recurrence.

Objective To explore the method of primary isolation, cultivation and identification of rat brain microvessel pericytes. Methods The brain tissue of 10 3 week-old Wistar rats was separated sterilely. The brain microvessel fragments were separated using two-step enzyme digestion and one-step gradient centrifugation and were seeded in 35-mm dishes for primary culture. The cell morphology was observed by phase contrast microscopy; the immunofluorescence assay was used to identify the associated antigns, such as the α-smooth muscle actin (α-SMA), neuron-glial antigen 2 (NG2), von Willebrand factor (vWF), and glial fibrillary acidic protein (GFAP). Methyl thiazolyl tetrazolium was used to determine the cell growth curve. Results Pericytes climbed out from the adherent brain microvascular fragments around,showing polygonal, and the cell fusion was 95％ after 12-14 days. Immunofluorescence staining revealed that the molecular markers of the pericytes α-SMA and NG2 related antigens showed double positive, while the vWF and GFAP related antigens showed double negative and the cultured cells were confirmed as brain microvascular pericytes. The growth rate of primary cells was slower. The passage cells entered into logarithmic growth phase after 36 to 60 hours and entered into plateau phase after 72 to 108 hours. Conclusions This method may successfully isolate rat brain microvascular pericytes with higher purity.

Purpose:To evaluate the feasibility, advantages and disadvantages of chemosensitivity test of human gastric cancer using MTT assay with short term culture tumor cells contaminated by nonmalignant cells compared with purifiedly primary culture tumor cells.Methods:Fifty nine fresh samples from patients with gastric cancer were obtained from operating rooms. Chemosensitivity results were provided by the MTT assay with short term culture cells. The primary culture cells were purified by means of a series of methods such as repeated cell attachment, differential trypsinization and natural purification which removed fibroblasts and other nonmalignant cells. The same MTT assay was conducted using purified cells. Chemosensitivity results between short term method and purification method were compared for seven antitumor drugs. Results:The success rate of short term method using the MTT assay for chemosensitivity testing was 81 4% (48 of 59 patients),and for the purification method it was 50.8(30 of 59 patients). The average was 20.2?9.5 days when primary cells were cultured into purified cancer cells. The optical density ( A ) value correlated directly with the the number of tumor cells with the two methods( P

Objective To study the feasibility of high performance liquid chromatography-evaporative light scattering detector ( HPLC-ELSD) in quantitative analysis of multi-components ( QAMS) by single marker. Methods Four saponins in Astragali Radix were simultaneously determined by HPLC-ELSD using external standard method, and malonylastragaloside I served as internal standard. The relative correction factors between internal standard and astragaloside Ⅰ, astragaloside Ⅱ and astragaloside Ⅳ were calculated, and the stability was investigated. Results Astragaloside Ⅳ in Astragali Radix was little, while malonylastragalosideⅠand AstragalosideⅠwere abundant.The relative correction factors lacked stability, so ELSD could not be used in QAMS. Conclusion HPLC-ELSD can precisely determine contents of four saponins in Astragali Radix. The detector needs to be further studied when the components have poor ultraviolet absorption such as saponins by QAMS.

Objective To study the expression of the excision repair cross complementing 1 ( ERCC1 ) and thymidylate synthase ( TS) in gastric cancer and to investigate the correlation between the ERCC 1 and TS ex-pression with the efficacy of FOLFOX adjuvant chemotherapy in gastric cancer patients .Methods A total of 73 gastric cancer patients received modified FOLFOX adjuvant chemotherapy .Real-time PCR was used to investi-gate the expression of ERCC 1 and TS in gastric cancer tissue .We also studied their correlation and association with prognosis .Results ERCC1 and TS expressed in gastric cancer tissues with the positive rate were 42 .5%(31/73)and 61.6%(45/73).There was no significant correlation between the ERCC 1 or TS expression and the clinical features .In this study ,the median of RFS and MST were 16 months and 27 months in patients with ER-CC1-negtive expression,9 months and 16months in patients with ERCC1-positive expression,the median of RFS and MST were significantly different in the two groups (P=0.000,P=0.002).There were no statistical differences on TS expression pattern in either median of RFS or MST (P=0.10,P=0.71).Conclusion The ex-pression of ERCC1 may be useful in prediction of the clinical outcome in gastric cancer patients who treated with FOLFOX7 adjuvant chemotherapy .Patients with ERCC1-negtive expression may benefit from the therapy .