Objective To establish the fingerprints for characterization of the chemical components of maidong(root of Ophiopogon japonicus) in two main cultivate regions of China,Sichuan(Chuanmaidong) and Zhejiang(Hangmaidong).Methods An HPLC-UV analytical methods was applied to detect 70% ethanol extracts of 20 samples from Sichuan and Zhejiang province,a "Fingerprint similarity evaluating system for TCM" issued by Pharmacopoeia Committee of P.R.China was used to evaluate the similarities all of the samples.Results The fingerprints revealed that the similarities were higher than 0.95 between samples from the same cultivate region,and were lower than 0.80 between samples from different regions of above two.Conclusion The fingerprints of Chuanmaidong and Hangmaidong were provided with high difference,and the difference can be taken as a most important proof for distinguishing the material medica of Chuanmaidong and Hangmaidong,but also in patent medicine of TCM.

Objective To evaluate the feasibility of transfection of Sonic hedgehog gene (SHH)into bone marrow mesenchymal stem cells(BMMSC).Methods After the SHH gene was transfected into BMMSC by electroporation apparatus,the transfection rate was evaluated by fluorescence inverted microscope.The growth curves of untransfected and transfected BMMSC were drawn,respectively,to observe the influence of transfection on cells.The expression of SHH gene in the BMMSC was detected by PCR,RT-PCR,Western-blot analyses.Results Through fluorescence inverted microscope,the observed transfection rate was appropriately 30％,PCR showed a obvious increase of SHH expression in transfected cells than that in untransfected cells,and it is quantified by qPCR for appropriately 7 times.Western-blot further demonstrated that the SHH protein expression in transfected cells had a distinct increase.However,it was observed that the exponential phase of BMMSCSHH growth curve delayed.The growth curves of both overlap 12 days after transfection.Conclusions This electroporation method can transfect exogenous SHH gene into BMMSC sufficiently with the effective protein expression in BMMSCSHH.It is the foundation of further research of genetic therapy for ischemic heart disease.

Objective To investigate the survival and differentiated situation of transplanted mesenchymal stem cells ( MSCs) transfected with Sonic hedgehog (SHH) gene after chronic myocardial infarction (MI).Methods MSCs were obtained from bone marrow of limb bones by density gradient centrifugation combined with attachment culture method .SHH gene was cloned and eukaryotic expression plasmid pcDNA 3.1-SHH was constructed .Then nucleofector TM was used to transfer SHH gene into MSCs and the expres-sions of mRNA and protein of SHH gene were detected in vitro .Myocardial infarction model was established by ligating the left anterior descending branch .A total of 150μl (3 ×106 ) of MSCs transferred with SHH gene was injected into ischemic area at the fourth week after MI model was set up .Brdu immunohistochemical staining and electron microscope examination were used to detect the survival and differentiation of MSCs with SHH in vivo at the 1st, 2nd, 4th, and 8th week after MSCs SHH transplantation.Results It was con-firmed that SHH gene clone and the recombinant eukaryotic expression vector pcDNA 3.1-SHH construction were successful by gene se-quence analysis and double digestions with EcoR I and Bamh I .Reverse transcriptase polymerase chain reaction ( RT-PCR) and West-ern-blot demonstrated that the expression of SHH mRNA was significantly increased in transfected MSCs compared with untransfected MSCs.Brdu immunohistochemical staining confirmed MSCs with SHH were survived from one to eight weeks in MI area after transplan -tation.Electron microscope examination showed that MSCs had the trend to be differentiated into myocardiac -like cells in MI area at the 4th and 8th week after transplantation .Conclusions The expression of SHH mRNA was significantly increased while MSCs were transfected with SHH gene , and MSCs SHH could survive and differentiate in good condition in the transplanted recipients .Those data provide the experimental basis for further transgenic cell therapy research of ischemic heart disease .

Objective To study the dosimetry and safety of the non-coplanar IMRT plan for advanced lung cancer.Methods The two groups IMRT plans were designed with coplanar (5,7F) and non-coplanar field (5,7F-n) for patients.To compare the dosimetry of two groups and perform 4 patients F7-n IMRT plan.Results With the increase of the fields in each group PTV's CI were improved (all P =0.000),especially the 7F-n plan PTV's Dmean,Dmax,V95％ and HI also were improved (P=0.001,0.001,0.009,0.000) ; in the coplanar group each lung' s V5 increased (P =0.000,0.002,0.000) and whole lung's Dmean increased (P =0.000),but non-coplanar group whole lung's and contralateral lung's V5 reduce (P =0.001,0.005).Between the groups,7F-n plan PTV's indicators were all improved to compared with 5F plan (all P =0.000),and each lung's V20 reduced (all P =0.000),and whole lung's Dmean,V30,contralateral lung' s V5 reduced (P =0.000,0.001,0.000),and spinal cord' s Dmax also reduced (P =0.033),but ipsilateral lung's V5 and heart's Dmean increased (P =0.000,0.003);with compared to 7F plan,the 7F-n's ipsilateral lung's V5 and heart's Dmean also increased (P =0.000,0.048),but whole lung' s and contralateral lung's V5 decreased (all P =0.000).Four patients were performed successfully non-coplanar IMRT treatment,no collision occurred.Conclusions 7 fields non-coplanar IMRT plan not only improve the dose distribution of PTV,but also effectively control the volume of low dose lung increase,lung V20 and Dmean reduce too.Thus recommended to use this design in patients with advanced lung cancer for radiotherapy

Objective To investigate the effect of heme oxygenase-1 ( HO-1 ) on the acute radiation-induced skin injury by gene transfer.Methods Thirty-three male SD rats were randomly divided into three groups as PBS-injected group,Ad-EGFP-injeeted group and Ad-HO-1-injected group ( n =11 ).In each group,three rats were used for determining the expression of target gene and the other rats were irradiated on the buttock skin with 40 Gy electron beam generated by a linear accelerator.Immediately after irradiation,rats were administered with a subcutaneous injection of PBS,Ad-EGFP or Ad-HO-1,respectively.Subsequently,the skin reactions were measured twice a week using the semi-quantitative skin injury scale.Results The strong positive expression of HO-1 was observed in subcutaneous dermal tissue after injection of Ad-HO-1.Compared to the PBS-injected group or the Ad-EGFP-injected group,a significant mitigation of skin injury was observed in Ad-HO-1-injected mice 14 d after irradiation (q =0.000-0.030,P ＜ 0.05 ).Conclusions HO-1 could significantly mitigate radiation-induced acute skin injury and Ad-HO-1 could be used to treat radiation-induced skin injury.

This study was aimed to explore differential effects of various sections of the velvet antler on promoting cell proliferation in vitro. The NRK-49F cell line from rat kidney fibroblasts was used as the cell model. The cell proliferation rates of the water extracts from the upper, middle and lower section of fresh velvet antler were measured by the MTT method. BCA method was used in the detection of protein concentration. The SDS-PAGE method was used in the analysis of difference composition of the sample protein. The results showed that soluble protein content of the upper, middle and lower section were 17.89, 16.04 and 6.89 mg·mL-1, respectively. From the top to the base, the soluble protein content of velvet antler was decreased. After 24 h treatment, when the protein concentration of the upper and middle section samples of the velvet antler were 800 μg·mL-1 and 600 μg·mL-1, the cell proliferation promoting rates reached the maximum, which were 66.76% and 64.36%, respectively. And when the lower section sample of the velvet antler was 1 000 μg·mL-1, the cell proliferation promoting rates reached the maximum, which was 58.87%. After 48 h treatment, when the upper and middle section samples of the velvet antler were 800μg·mL-1, the cell proliferation promoting rates reached the maximum, which were 219.56% and 215.86%, respectively. And when the lower section sample of the velvet antler was 1 000 μg·mL-1, the cell proliferation promoting rates reached the maximum, which was 169.20%. The velvet antler on the proliferation of cells was much better than the 10% fetal bovine serum. The figure of SDS-PAGE showed the slight difference in the protein composition of three part of the velvet antler. It was concluded that all samples had promoting effects on cell proliferation with concentration-depen-dent, and the main protein in different part of the velvet antler had minor differences.

Objective To compare clinicopathological characteristics and prognosis between triple-negative and HER-2-overexpressed breast cancer patients.Methods From Jan.1997 to Jan.2007,data of 725 cases of primary breast cancer patients undergoing surgical treatment were analyzed.The patients were classified into triple-negative and HER-2-overexpressed phenotypes based on immunohistochemistry results.The clinicopathological characteristics and survival of the 2 groups were compared.Results Of the 725 cases,triple-negative and HER-2-overexpressed phenotypes accounted for 12.29%and 24.96%respectively.Compared with HER-2-overexpressed group,the triple-negative group had significantly higher proportion of familial history of malignancy (18.4% vs 5.5%,P=0.001)and higher proportion of patients'histological grade reaching grade 3(54.0% vs 42.0%,P=0.01).There were more lymph node metastasis in triple-negative group than that in HER-2-overexpression group(N1+N2+N3:74.7%vs 64.6%,P=0.045).The recurrence and metastasis rate within 2 years and brain metastasis rate in triple-negative group were higher than those in HER-2-overexpressed group (25.3%,8.0%vs 8.8%,2.2%,respectively,P<0.05).The 5-year disease-free survival rate in triple-negative group was significantly lower than that in HER-2-overexpressed group(55.6%vs 69.8%,P=0.041).There was no statistical difference between the 2 groups in terms of age,menopausal status,tumor size,pathological stage,surgcal procedure,pathological type,adjuvant radio-chemotherapy,the proportion of metastasis to liver,lung and bone as well as overall survival rate(P>0.05).Conclusion Compared to HER-2-overexpressed group,patients with triple-negative breast cancer show higher rate of familial history of malignancy,and they are associated with higher histological grade and poor prognosis.

DNA polymerase iota (Polt),as well as Revl,Polκ and Polη,are all Y family DNA polymerases,which are able to replicate damaged DNA via translesion synthesis pathway.However,Pol(t) has the lowest fidelity among all DNA polymerases in both correct and inaccurate DNA templates.Also Pol(t) can bypass certain DNA damages and accumulate mutations.Recent studies show that the aberrant expression of Pol(t) is observed in human uveal melanoma,breast cancer,bladder cancer,lung cancer and esophageal cancer,which may contribute to the tumorigenesis and progression of tumor.The special role of Pol(t) in replicating damaged DNA may contribute to the resistance in oncotherapy.

Objective To investigate the differences of clinicopathologic characteristics and prognosis between young and old age patients with rectal cancer. Methods From January 1996 to January 2006, 85 young patients(age≤40 years) and 155 older patients(age≥65 years)with rectal cancer were surgically treated. The clinicopathological and follow-up data of them were retrospectively analyzed and compared by survival analysis and COX regression multivariate analysis. Results Rectal cancer under peritoneal in young group were higher than that in older group (69.41 % vs 52.90 %, P =0.013). The young group had significantly higher frequencies of pooly differentiated carcinoma (31.76 % vs 18.71 %, P =0.023) and more mucinous adenocarcinoma as well as signet-ring cell carcinoma (22.35 % vs 8.39 %, P =0.007), There were more lymphatic metastasis in young group than that in old group (N_1+N_2: 63.53 % vs 47.10 %, P =0.015). The overall 5-year survival rates were 48.2 % and 55.7 % in young and old patients respectively, which was not significantly different (P =0.176). COX regression showed that radical operation, tumor infiltration depth,lymph node metastasis and TNM stage were independent prognostic factors. Conclusion As compared to the old age patients, more malignancy and more advanced stage are common in young patients with rectal cancer.However the efficacy of young patients is similar to the older counters by early detection and radical operation combined radiotherapy as well as chemotherapy.

Some of the patients with rectal cancer are less sensitive to preoperative concurrent chemoradiotherapy (CCRT).Patients who are resistant to CCRT have a poor local tumor control and CCRT may also increase adverse reactions.The sensitivity of rectal cancer patients to CCRT can be predicted by magnetic resonance imaging (MRI),positron emission tomography,serum carcinoembryonic antigen,molecular biomarkers and gene expression profiling before treatment.According to the predicted results,the clinicians are instructed to choose individualized treatment for the patients so that the therapeutic effects of rectal cancer are further improved.