Objective To explore the effects of specific T-cell immunity against prostate cancer(PC) cells induced by dendritic cells(DCs) derived from fetal organs.Methods Mononuclear cells(MNCs) were obtained from the fetal bone marrow and liver.Then MNCs were cultured in medium with induction of rhGM-CSF,rhIL-4 and rhTNF-?to get DCs.Lysates of DU145 containing HSP-peptide complex were prepared by 50%-70%(NH4)2SO4 saturation.T lymphocytes from fetal spleen were co-cultured with DC loading DU145 antigen for 72 h,whereby CTL was obtained.The cytotoxicity of CTL against DU145,PC3 and EJ was detected by MTT assay.Results Mature DCs were induced from fetal organs,which expressed CD1a,CD_(86),HLA-DR and CD_(83) at high levels.DC stimulated with tumor lysates transformed T cells to specific CD~+_8 CTL.Phenotype of CD~+_8 cell was(14.09?(2.46))% before transformation,and(62.76?2.64)% after transformation,respectively(P

According to the existing Provisions for Drug Registration (SFDA Order No. 28), applications for new drugs of traditional Chinese medicine are divided into two parts: the applications for drug clinical trial and for drug production (including new drug certificate). It will last for about 10 years from the application for drug clinical trial to get approving, and it also remains many problems and the low probability to succeed. From the sight of pharmaceutical review, there are mainly two aspects of regulatory compliance and technical issues, mainly for changes without approval of the competent authorities of the country. For example, sample preparation and approval of clinical trial process are significant changes. Technical problems are reporting incomplete data or information submitted does not comply with the technical requirements for review, such as: production process validation does not provide information, the preparation of samples for clinical trials and field inspection, production information, or the information provided does not meet the technical requirements. This paper summarizes the frequently asked questions and to make recommendations to advise applicants concerned, timely detection of problems, avoid risk, improving the quality and efficiency of the application for registration.
China ; Drug Approval ; legislation & jurisprudence ; Drug Evaluation ; legislation & jurisprudence ; Humans ; Legislation, Drug ; Medicine, Chinese Traditional

The aim of this study was to evaluate the effects of compound probiotics on milk performance,blood biochemical indexes,rumen fermentation parameters and nutrient digestibility of Holstein dairy cows in middle-and-later lactation.Forty-eight dairy cows with similar milk yield,days in milk and age were randomly divided into 4 groups treated with four different levels of compound probiotics as follows:the control group (0 g/d),group 1 (10 g/d),group 2 (20 g/d) and group 3 (30 g/d).The adaptation period was 7 days,and the total experimental period was 60 days.Results showed that:compared with the control group,supplementation of compound probiotics could improve milk yield and significantly increase the concentration of milk protein,lactose and total solids (P＜0.05);there was no significant effect on blood biochemical indexes (P＞ 0.05);compound probiotics could significantly increase the content of ammonia and microbial protein content (MCP) in the rumen (P＜ 0.05);the apparent nutrient digestibility of crude protein and crude fat of group 2 was significantly higher than other groups (P＜0.05);compared with the control group,the economic benefits of group 1,2,3 increased 2.28,4.80 and 4.09 yuan/d,respectively.In summary,dietary supplementation with 20 g/d of compound probiotics was the most effective method for milk performance,rumen fermentation parameters,nutrients apparent digestibility and economic benefit of Holstein dairy cows.

OBJECTIVE To investigate the positive rate of ?-lactamase,oprD2,aminoglycosides modifying enzyme and qacE△1 gene among clinical isolated strains of multi-resistant Pseudomonas aeruginosa in our hospital.METHODS P.aeruginosa was determined by VITEK,and MIC was determined by agar dilution method.TEM,IMP,VIM,oprD2,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6″)-Ⅱ,ant(3″)-Ⅰ,ant(2″)Ⅰ and qacE△1 in strains were detected by polymerase chain reaction(PCR).RESULTS The positive rate of TEM,aac(3)Ⅱ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰ in multi-resistant P.aeruginosa were 51.4%,48.6%,40.0%,54.3%,45.7% and 60.0%,respectively.No IMP and VIM genes were detected.CONCLUSIONS Positive rate of ?-lactamase, aminoglycosides modifying enzyme is high in 35 tested strains,and that should be paid more attention.

ABSTRACT:Objective To a rare case of double primary cancer (hepatocellular carcinoma with gallbladder)to guide clinical application.Methods and Results We reported one case of primary hepatocellular carcinoma and gallbladder.A male patient,54 years old,had main complaints of intermittent right upper quadrant pain for 4 days. The abdominal CT of the local county hospital showed gallstones,gallbladder with liver infiltration.And then he went to the First Affiliated Hospital of Xi’an Jiaotong University for further treatment.Laboratory examination revealed:HBsAg(+),HBcAb(+),alpha-fetoprotein (AFP)> 60 500 ng/mL,carcinoembryonic antigen (CEA) 5.25 ng/mL.Abdominal CT showed hilar slightly stronger light echo groups:liver cancer or gallbladder cancer? Hepatic artery and portal vein CT imaging (CTA+CTV)examination showed the malignant tumor shadow in the inside of the left hepatic lobe huge,uneven thickening of the gallbladder wall,suspected liver disease with gallbladder infringement or gallbladder disease with liver infringement. With the preoperative preliminary consideration of primary liver cancer with infiltration of the gallbladder,we chose the operation as the resection of segment Ⅳ b and Ⅴ of the liver,cholecystectomy and T tube drainage.Pathological examination postoperative showed the bulky liver carcinoma grade Ⅲ and the poorly differentiated adenocarcinoma of gallbladder.A month later,abdominal CT showed the tumor spread intrahepatic,prompting the poor prognosis.Conclusion The two which are not continuous,which is the standard of double primary cancer,are not suitable for all double primary cancers.This case provides useful experience for future similar diagnosis and treatment of disease,and also helps us with timely and accurate identification of “metastatic”or “primary”,which is the key point for clinicians to give patients an effective treatment.

Desensitization, Immunologic ; methods ; Humans ; Injections, Subcutaneous

Adult ; Amniotic Fluid ; physiology ; Female ; Humans ; Kidney ; abnormalities ; diagnostic imaging ; Pregnancy ; Retrospective Studies ; Ultrasonography, Prenatal

Objective To construct an expression plasmid carrying the specific siRNA of survivin gene,and to evaluate its silencing effect on the expression of survivin gene and its inhibition effect on the growth of gastric cancer cells.Methods The specific siRNA of survivin gene was designed and synthesized,and an expression plasmid pAdGFP-siRNA was constructed.Gastric cancer cell line SGC-7901 was cuhured and transferred with pAdGFP-siRNA,then the silencing of survivin gene expression and the growth inhibition of cancer cell mediated by pAdGFP-siRNA were identified.Results The growth of gastric cancer cells was inhibited after transferring the pAdGFP-siRNA,with the inhibition rate of 68.2% compared to the control group.Immunohistochemistry showed that the specific siRNA markedly silenced the expression of survivin gene in cancer cells.Conclusions The overexpression of survivin gene in gastric cancer cells results in the high proliferation and the resistance to the chemo- and radio-therapy of the cancer cells.The specific siRNA can markedly silence the expression of survivin gene and inhibit the growth of cancer cells.

OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.

METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.

RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.

CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.

Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection

Objective Our hypothesis was if we rendered host' s cytotoxic tumor lymphocytes insensitive to TGF-β,these immune cells could be able to overcome the TGF-β mediated immunosuppression and reject the tumor.We aimed to develop a lentivirus mediated gene transfer program incorporating a herpes simplex virus thymidine kinase(HSV-tk)in our dominant negative TGF-β type Ⅱ receptor(TβRⅡDNglytk)expression vector.So we first need to construct the lentiviral pLenti6/V5-D-TOPO vector containing TβRⅡDNglytk and produce the recombinant lentivirus as the transfection vector.Methods PCR were used to amplify the genes TβRⅡDN and HSV-tk from the respective plasmids.Then the genes were linked by recombinant PCR technology to construct the fusion gene TβRⅡDNglytk and control vector TRANSglytk.According to the operation manual from the Invitrogen company,TOPO cloning technology was used to construct the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk.Both of the constructed plasmids were verified by sequencing.ViraPowerTM Lentiviral System and 293 FT cells provided by Invitrogen were used to produce the recombinant lentivirus vector,the tillers of the lentivirus were determined by 293 cells.Results The construction of the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk were completed succefully.The DNA sequencing results showed that both the plasmids were constructed correctly.Using them we successfully produced infectious lentivirus vectors with appropriate tilters.Conclusions Using TOPO cloning technology in the construction of the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk and recombinant PCR in the fusion genes is feasible.Recombinant PCR combine with TOPO cloning technology can be a simple,highly efficient and rapid way to construct lentiviral vector.The production of infectious lentivirus with appropriate tilters using 293FT is suitable and feasible.The construction of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and production of the infectious lentivirus will lay a foundation for immunotherapy of prostate cancer.