Humans ; Integrative Medicine ; Medicine, Chinese Traditional

Shengmai Injection (SMI), is a traditional Chinese medicine (TCM) formula injection composed of Panax ginseng, Ophiopogon japonicas and Schisandra chinensis, which is widely used in clinic for the treatment of cardiovascular diseases as well as protecting against pulmonary disease and add-on therapy to cancer chemotherapy. In recent years, the pre-clinical basic research of SMI was widely performed. The studies, including chemical composition, in vivo process, and action mechanism and so on of SMI, helped to provide scientific foundation for revealing the material basis of formula. In this paper, the studies on material composition, pre-clinical pharmacokinetics and pharmacodynamics of this formula were summarized, so as to provide reference for the quality control, second development and rational clinical application of SMI.

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Objective To lay the physiological and biochemical basis for establishing and evaluating tree shrews model of human disease. Methods There were 92 Tupaia belangeri chinensis, in which half of them were male,they were allowed to eat nothing for 12 hours, then we sampled heart blood 0.8~1.0 mL without anesthesia and put blood samples into sterilized centrifuge tube for separation and preparation of serum. Olympus AU5400 automatic biochemistry analyzer was used to measure biochemical indexes. Then 1.5~2.0 mL of urine of each tree shrew was collected and put into sterilized centrifuge tube for measuring renal function by using Combi-scan500 urine analyzer. Fianally SPSS statistics software was used to analyse the measured values, and comapared the measured values with the human reference values. Results There were significant differences in myocardial enzymes and some renal function indexes such as lactic dehydrogenase,α- hydroxybutyric, acid dehydrogenase, creatinine, uric acid, urine specific gravity and pH value between male and female Tupaia belangeri chinensis ( <0.05), and there was no significant difference in the rest of the measured values ( >0.05) . Then determination value of Tupaia belangeri chinensis, male’s and female’s, myocardial enzyme and part of the renal function indexes were compared with the human reference values. Some indexes including urea, urine specific gravity, urine, urobilinogen were in the range of human reference value, while the values of urine bilirubin, urine nitrite. lactate dehydrogenase, creatine kinase, α-hydroxybutyrate dehydrogenase, creatine kinase isoenzyme CKMB were higher than the human reference value. White blood cell, urine protein, ketone, occult blood most of them were negative as the same to human reference value, but sometimes were positive, and the positive rates respectively were 3.95%and 46.1%,39.5%,2.63%. The measured value of Vitamin C was positive that is completely opposite to human reference value, but sometimes is negative, the negative rate was 6.6%. Conclusion Urea, urine specific gravity, urine, urobilinogen, urine bilirubin, urine nitrite can be directly used as the indexes for evaluating tree shrews models of human disease, other indexes can be used as indexes for judgment of the normal physiological and biochemical basis of tree shrews.

Based on the gene sequence of AIV matrix protein 2(M2) and two cytotoxic T-lymphocyte epitopes derived from nucleoprotein,the prokaryotic expression vector pET-3M2e-NP1-NP2 was constructed.The target gene was expressed in the solvable form in BL21(DE3) when induced with 1.0 mmol/L IPTG and Western blot analysis showed that the expression product had good immunogencity.Purified fusion protein was mixed with various amount of adjuvant,including Freund's,Vash oil and chitosan,and then immunized 20-day-old chicken by intramuscular injection and boosted 3weeks later.Blood samples were collected weekly following the primary vaccination.The anti-M2e antibody was detected with ELISA method with the synthesized peptide as coated antigen;the neutralizing ability of anti-serum was evaluated on MDCK cell line and chick embryo,the CD4+ and CD8+ T lymphocyte amounts in peripheral blood of immunized chicken was measured by flow cytometry.Results showed that the fusion protein can induce immunological reaction,and the antibody can bind with the viral M2 protein that expressed on the surface of MDCK cells.Flow cytometry result showed that CD4+ and CD8+ T lymphocyte in peripheral blood increased obviously following immunization(P

M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.

Objective To construct an eukaryotic expression vector with human adipose most abundant gene transcript 1 (APM1) gene,and to investigate the transfection and expression of pCDEF-APM1 eukaryotic expression plasmid in HEK293 cells.Methods pCDEF-APM1 eukaryotic expression plasmid was constructed by DNA recombinant method.Expression vector pCDEF-APM1 was transfected into HEK293 cells with Effectene reagent.The level of human adiponectin protein in the supernatant of cell culture media was detected with double antibody sandwich ELISA.Results The sequence of DNA fragment from constructed pCDEF-APM1 plasmid was identical to that published in GenBank.There was raised human adiponectin protein level in culture supernatant of HEK293 cells tnmsfected with pCDEF-APM1.Conclusion The pCDEF-APM1,an eukaryotic expression plasmid for APM1 gene is successfully constructed.High protein expression of adiponectin can be obtained in HEK293 cells transfected with pCDEF-APM1 eukaryotic expression plasmid.

Objective To evaluate the clinical value of 18floro-deoxyglucose positron emission tomography-CY(~(18)FDG PET-CT)in the diagnosis of lymph node metastasis from advanced esophageal carcinoma. Methods A prospective study is perfonued here to assess whether ~(18)FDG PET-CT can improve the diagnostic accuracy in lymph node metastasis for patients with advanced esophageal carcinoma.Thirty patients had undergone esophagectomy with extensive lymph node dissection.PET-CT findings were compared with that d CT with pathological finding as the final say.Results All patients were operated successfully without peri-operative complications.The pathological examination conformed metastasis in 22 patients and 49 out of 243 excised lymph nodes.In CT analysis,the sensitivity was 40.8%,specificity was 96.9%,with a diagnostic accuracy of 85.6%, The positive and negative predictive value was 76.9%,86.4% respectively;PET-CT resulted in a sensitivity of 93.9%,specificity of 91.2%,accuracy of 91.8%.The positive predictive value was 73.0% and negative predictive value was 98.3%,The difference of sensitivity(P＜0.001),accuracy(P＜0.05)and negative predictive value between the two radiological modalities was statistically significant(P＜0.001).Conclusions With a high sensitivity and accuracy in the diagnosis of lymph node metastasis,PET-CT appears necessary in preoperative examination for advanced esophageal carcinoma in the hope that surgical treatment be guided by the results of PET-CT,especially for the elder patients with poor pulmonary function or heart or brain complications. Moreover,it could be used as the basis of the conformal radiation therapy planning for inoperable patients.

Objective To evaluate the clinical value of 18-fluoro-deoxyglucose positron emission tomography-CT(~(18)FDG PET-CT) for recurrence and metastasis in treated esophageal carcinoma (EC). Methods A retrospective study is done on 37 previously treated EC patients who underwent PET-CT scans to detect recurrent or metastatic lesions.The diagnostic accuracy of ~(18)FDG PET-CT was assessed with the help of pathological finding as well as clinical or follow-up data.Results Fourty-six sites of recurrence were finally confirmed in 37 patients by cytology,pathology or follow-up data.The sensitivity,specificity and accuracy of PET-CT in detecting recurrence of all sites were 93.5% (43/46),76.9% (20/26) and 87.5% (63/72),respectively.Two false-positive findings were found both at the anastomosis and hilar nodes,which caused the decrease in the overall specificity,especially that locally.The analysis of standard uptake value (SUV) demonstrated that patients with recurrence or who died during follow-up had higher SU- Vs compared with the control group.Condusions ~(18)FDG PET-CT is highly effective in detecting recur- rence in previously treated EC patients despite the low specificity at local sites.The analysis of stardard up- take value(SUV) provides incremental value in prognosis for this patient cohart.

Objective To determine the effect of nanochemotherapy drug on Survivin and p53 ex-pressed by human biliary traet carcinoma cell line QBC939.Methods Culturing the human biliary tract carcinoma cell line QBC939 in vitro and it was divided into five groups including saline,nanochemotherapy drug,nanopartiele withoul nanochemotherapy drug,5-FU and gemcitabine.Using the methods of MTT and flow cytometry to observe the growth of QBC939 which was dealt with different drugs.In addition,RT-PCR and Western blot were used to delect the expression of mRNA and protein by Survivin or p53.Results The expression of mRNA and protein by Survivin decreased in the following set:saline,nanoparlicle withoul nanochemotherapy drug,5-FU,gemcitabine and nanochemotherapy drug,respeclively.However,the ex-pression of mRNA and protein by p53 were in reverse order.The inhibiting action to QBC939 was obvious in nanochenmtherapy drng and the apoplotic rate was higher than others except for gemcitabine(P＜0.05). Conclusion Nanochemotherapy drug has significant effect on treatment cholangiocarcinoma in vilro,which may attribute to the down regulation of Survivin and up regulation of p53.