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 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 .

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 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 study cervical HPV infection in female patients with vulvar condyloma acuminatum(CA)from Shanghai area.MethodsExfoliated cells were obtained from cervices and vulva lesions of CA of 194 patients.respectively.HPV genotyping was carried out in cell samples using capture-hybridization method and gene chip techniques.Results HPV was detected in vulva lesions of all the 194 patients.Among them,74.2%(144/194)were positive for low risk(LR)-HPV,and 25.8%(50/194)for high risk(HR)-HPV.A single HPV genotype(6 or 11)was detected in 136(94.4%)patients with LR-HPV.and mixed genotypes of LR-HPV and HR-HPV in 46(86%)patients with HR.HPV.Of the 194 patients.85.6%(166/194)were complicated by cervical HPV infection.including 119(61.4%)cases of LR-HPV and 46(23.7%)cases of HR-HPV,In the case of HPV genotype.the consistence between CA lesions and cervix was 95.8%(159/166).The prevalence of LR-HPV declined sequentially from subtype 11 to 6 and 53,and that of HR-HPV from subtype 16 to 18 followed by 52,31,45 and 58.Conclusions There is a high rate of infection with HR-HPV in female patients with CA.and nearly one quarter of these patients are complicated by cervical HR-HPV infection.

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.

Objective To establish the athrmic inouse model of breast cancer in normal position and imitated metastatic breast cancer. Methods Breast cancer cells MDA-MB-231-luc carrying luciferase gene was injected into the athymie mice.The optieal imaging in vivo system was used to observe the establislament of the model. Reseults The breast tumor emerged after we planted the MDA-MB-231-lue cells in the mammary gland fatpads,the volume and photon of the tumor increased during the second weekto the fifth week.After injection by the tail vein,the tumors mainly located in the lungs While after infusion in the left alrtrium.the tumolrs metastate to all over the body. Conclusions We succeeded in the establishment of the athymic mice model of breast cancer.in situ and imitated metastatic breast cancer by iniection into the vena caudalis and the left alttrium.The optical imaging in vivo system could distinctly show the formation of the tumors.