Hoxa3-Pax1/Pax9-Eya1-Six1/4 regulatory pathway seems to be operating during forming the bilateral parathyroid/thymus common primordial in early embryonic development.The specification of the parathyroid domain in the parathyroid/thymus primordial is regulated through a Shh-Tbx1-Gcm2 pathway.Gcm2 also may play roles in later steps of parathyroid development,including CaSR and PTH gene expression.MafB and Gcm2 interact with each other and synergistically activate PTH transcription.Genetic basis and the etiology of some hypoparathyroid disorders in man are involved defects in transcription factors that include GCMB,GATA3,Tbxl,SOX3 and GNA11.This marker expression in thymus and parathyroid primordium includes HoxA3,Pax1,Eya1,and Six1;and expression of parathyroid cell-like cells includes Gcm2,CaSR,and PTH.These expressions may serve as markers of stem cell differentiation into parathyroid cell-like cells.

[Objective] To investigate the expression of MICA mRNA and protein in osteosarcoma and give a more comprehensive understanding to its immune evasion.[Methods] RT-PCR was used to analyze MICA mRNA in 11 osteosarcoma tissues and 5 osteosarcoma cell lines.MICA expression was examined in 66 paraffin-embedded osteosarcoma tissues and 6 paraffin-embedded normal bone tissues by immunohistochemistry,and MICA protein in 9 fresh osteosarcoma tissues was detected by Western blot too.MICA surface expression was estimated by flow cytometry.[Results] Nine of eleven (81.8%) osteosarcoma specimens and all of the five cell lines consistently expressed MICA mRNA.Up-regulation of MICA expression was found in osteosarcoma (34/66,51.6%),compared with normal bone tissues (0/6,0%).The cell lines Saos-2,MG63,and HOS showed positive surface MICA expression,while the U2OS and OS732 showed very limited expression.[Conclusion] MICA mRNA and protein predominantly expressed on osteosarcoma tissues and cell lines.

In order to further study mouse embryonic stem cells(ES cells),lentiviral vector PLL-IRES-Nanog-Neo was constructed.Mouse ES cells overexpressed nanog by mediation of lentiviral were cultured on mouse fetal fibroblast feeders after 2 weeks under G418 media and examined according to gowth characteristics. Results were showed that 918 bp nanog fragments were expressed in mouse ES cells mediated by lentiviral vector PLL-IRES-Nanog-Neo,mouse nanog-ES cells were taken on mass-like image and positve with alkaline phosphatase staining and Oct4 and SSEA1 immunocytochemistry under no LIF condition in the media. It is concluded that mouse ES cells Elevated nanog gene expression by mediation of lentiviral were constucted and cultured.

OBJECTIVE To strictly manage the whole process for cleaning the laminar air flow(LAF) operating center to achieve the modernized hospital environment′s standard requests.METHODS Right before the use of the LAF operating center,scientific management was conducted strictly according the regulations and standards issued by state Ministry of Health.RESULTS Fulfilling the standards of the process for cleaning operating center was all for the goal of improving the efficiency of management.CONCLUSIONS A management is made efficient from checking all the things before use,monitoring them,and training people so that they can efficiently carry out their tasks.The purpose of the scientific management is to reach the expected quality.

Objective To investigate the efficacy and toxicity of neoadjuvant regional arterial chemotherapy in the treatment of advanced gastric cancer. Methods The clinical data of 158 patients with advanced gastric cancer and with the same clinical stages who were admitted to Renji Hospital of Shanghai Jiaotong University from February 2002 to May 2005 were retrospectively analysed. Preoperative regional arterial chemotherapy was applied to 76 patients (test group) and the remaining 82 patients only received surgical treatment (control group). The chemotherapy regimen was epirubicin (50 mg/m2) + cisplatin (60 mg/m2) + 5-fluorouracil (1000 mg/m2).This regimen was modified to oxaliplatin (130 mg/m2) + 5-fluorouracil (1000 mg/m2) since 2003, and surgery was performed 6-11 days after the chemotherapy. All patients received postoperative intravenous chemotherapy.The clinical effects, radical resection rate, operative complications and long-term survival of the two treatment methods were evaluated. All data were analysed using the chi-square test and Kaplan-Meier analysis. Results The radical resection rate was significantly higher at 86% (65/76) in the test group compared with 71% (58/82)in the control group ( x2 = 5.01, P ＜ 0. 05 ). The toxicity of the chemotherapy in the test group was mild. The postoperative complication rate was 20% (15/76) in the test group and 16% (13/82) in the control group, with no significant difference between the two groups (x2 = 0.41, P＞0.05). The median survival time was 41 months in the test group and 23 months in the control group. The 5-year overall survival rate was higher in the test group (44.6%) than that in the control group (29.1%) (x2 =3.95, P＜0. 05). Conclusions Neoadjuvant regional arterial chemotherapy is well tolerated by patients with advanced gastric cancer. It is also effective for increasing the radical resection rate and improving the long-term survival.

To predict and identify the dominant B‐cell epitopes of conserved region of Treponema pallidum repeat protein F (TprFN ) and provide the basis for development of polyvalent epitope‐based syphilis vaccine ,the amino acid sequence of TprFN was obtained from GenBank and analyzed with comprehensive meta‐analysis Mobyle ,ABCpred and IEDB online software .The peptides containing predicted epitopes were artificially synthesized . To obtain and measure the titers of antibodies against TprFN ,New Zealand rabbits were immunized with recombinant protein TprFN expressed in E .coli and identified by Western blot (WB) .Sera from TprFN‐immunized rabbits ,syphilis patients ,and normal human and normal rabbits were used to deter‐mine the immunoreactivity and specificity of 7 predicted peptides of TpFN by indirect ELISA .Comprehensive meta‐analysis of online software showed that P1 (43‐62AA) ,P2(57‐71AA) ,P3(81‐88AA) ,P4(89‐103AA) ,P5(125‐138AA) ,P6(231‐251AA) and P7(268‐279AA) might be the B‐cell epitopes .A protein was expressed in a soluble form and identified as TpFN by WB .The ELISA indicated that P1 and P3 were active with TprFN‐immunized rabbit sera and syphilis patient sera but not with negative control sera .These results indicate that P1 and P3 are the potential dominant B‐cell epitopes .

Objective To investigate the preopera ti ve diagnosis and the surgical treatment of primary retroperitoneal tumor(PRT). Methods The clinical dat a of 71 patients with PRT were retrospectively analyzed including clinical manif estation, radiologic studies, pathologic examination and surgical procedures. Results There were 32 cases of benign tumor, 38 cases of malignant tumor and 1 case of borderline tumor. Of the 32 patients with benign tumor, 31 underwent complete surgical resection. Of the 38 patients with malignant tumor, 29 underwent complete resection. One patie nt with borderline tumor underwent total resection. The 5-year survival rate of benign PRT was 89.29%, the 5-year survival rate of malignant PRT was 20.80%. Four cases of recurrent benign PRT underwent complete resection. Ten cases of r ecurrent malignant PRT underwent complete resection and 3 underwent partial rese ction. ConclusionsRadiolog ical study is crucial for the diagnosis. Complete resection is the key for the t reatment of PRT.

Astrocytes maintain homeostasis of neuronal microenvironment, provide metabolic and trophic support to neurons and modulate neuronal responses to injury. Rotenone specifically inhibits mitochondrial complex I, and long exposure to rotenone may increase the risk for Parkinson's disease (PD) and cause Parkinsonism. However, little is known about the role of astrocytes in the process of rotenone-induced dopaminergic neuron injury. In order to investigate this issue, we used MN9D cells as a cell model of dopaminergic neurons and rotenone as a toxin to initiate mitochondrial deficiency. MN9D cells treated with the normal medium or astrocyte-conditioned medium (ACM) were exposed to different concentrations of rotenone for different time followed by cell viability measurement by MTT assay. Besides, various concentrations of ACM and temporally different treatments were devised to evaluate protective efficiency of ACM. Growth curve of cells in the normal medium or ACM was continuously assessed by cell counting for 8 d. The influence of rotenone and ACM on cellular oxidative stress was determined by DCFH-DA staining followed by flow cytometric analysis. Glutathione (GSH) content after treatment of ACM or rotenone was measured by GSH assay kit. Our results showed that rotenone decreased viability of MN9D cells in a dose-dependent manner and ACM treatment significantly attenuated rotenone toxicity at each concentration. No significant difference in growth rate was observed between the normal medium and ACM treatment. Four concentrations of ACM, namely 1/3ACM, 1/2ACM, 2/3ACM and pure ACM, all displayed protection, increasing cell viability to (124.15+/-0.79)%, (126.59+/-0.82) %, (125.84+/-0.61) % and (117.15+/-1.63) % of the cells exposed directly to rotenone, respectively. Treatment with ACM through the whole experiment except the initial 24 h, 24 h before or at the same time of rotenone addition all exerted protective effects, with cell viability being (110.11+/-2.52)%, (113.30+/-2.36) %, (114.42+/-2.00)% of the cells exposed directly to rotenone, respectively. Conversely, ACM treatment 12 h after rotenone addition had no protective effect, with cell viability being (102.54+/-1.36)% of the cells exposed directly to rotenone. Moreover, ACM treatment up-regulated GSH level in MN9D cells nearly twofold. Incubation with 100 nmol/L rotenone for 24 h depleted GSH level by nearly two thirds of the control, but ACM treatment mitigated the drop of GSH level, maintaining its content at (147.83+/-0.63)% of the control. Consistent with GSH change, rotenone administration resulted in a positive rate of 96.24% of DCF staining, implying a great extent of oxidative stress, whereas treatment with ACM reduced the extent of oxidative stress to a positive rate of 78.31%. Taken together, these findings suggest that astrocytes protect MN9D cells from oxidative stress caused by rotenone, and GSH partially accounts for the protection. Therefore, astrocytes may play a protective role in the process of PD.
Animals ; Astrocytes ; physiology ; Cells, Cultured ; Cytoprotection ; Glutathione ; analysis ; physiology ; Neurons ; drug effects ; metabolism ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Rotenone ; toxicity

OBJECTIVEIn oder to treat periodontitis by using tissue engineering and gene engineering technology, the article established an transient expression system of bone marrow stromal cells (BMSC) modified by osteoprotegerin (OPG) gene and detected its expression using eukaryotic secreted expression pSecTag2/B-OPG plasmid.

METHODSBy solation and culture of BMSC in vitro, the identified recombined plasmid was transiently transfected into BMSC by Lipofectamine 2000 and OPG expression in BMSC was determined by RT-PCR and Western blot in 6 weeks.

RESULTSThe fragments of the recombinant plasmid digested with Hind III, EcoR I and BamH I and examined by 10 g/L agarose electrophoresis, were consistent with predicted size. The sequence of OPG was identical to the sequence provided by GeneBank [gi:33878056]. OPG transcribing in BMSC was confirmed by RT-PCR and OPG sustainable expressing in BMSC was detected by Western blot in 39 days.

CONCLUSIONThe transiently expression system of BMSC modified by OPG gene was successfully established.

Humans ; Mesenchymal Stromal Cells ; Osteoprotegerin ; Tissue Engineering ; Transfection

AIMTo analyze and compare the constituents of the volatile oil of Pogostemon cablin collected from different regions of cultivation and harvesting times in order to evaluate the quality of Shipai Huoxiang and to expound the chemical intension of Pogostemon cablin.

METHODSThe combination of GC and MS.

RESULTSThe volatile oil compositions of Herba Pogostemonis collected from various of cultivation regions and harvesting times are obviously different. Based on the chemical differences of the volatile oil compositions, Pogostemon cablin is divided into two chemotypes, Pogostone-type and Patchouliol-type. The former was cultivated in Guangzhou and Gaoyao regions, locally named as "Shipai Huoxiang"; the latter was locally named as "Hainan Huoxiang", cultivated in Wuchuan, Suixi and Leizhou regions of Guangdong Province and Wanning region of Hainan Province. The Pogostone-type contains rich oxygenated components, especially pogostone in the volatile oil compositions and poor non-oxygenated composition with patchouliol. The above chemical data may be used as evaluation standard for the authentic Shipai Huoxiang. The Patchouliol-type contains similar quantities of oxygenated and non-oxygenated composition, especially rich patchouliol with poor pogostone in oxygenated compositions, rich delta-guaiene and alpha-guaiene in non-oxygenation compositions. The contents of volatile oil and their constituents were varied due to different harvesting time. The yields of pogostone and volatile oil of Shipai Huoxiang was higher in July. The quality of the samples collected in this month was better.

CONCLUSIONAccording to the volatile oil compositions, there are two chemotypes (Pogostone-type and Patchouliol-type) in Pogostemon cablin plant. These two chemotypes correspond to the genotypes of plastid matK gene and nuclear 18s rRNA gene by cluster analysis.

Ecosystem ; Environment ; Lamiaceae ; chemistry ; classification ; Oils, Volatile ; analysis ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; classification ; Quality Control ; Seasons