Tumor was one of the most threatening disease of the human beings. Tumor marker plays a very important role in tumor diagnosis, estimating curative effects and prognosis. Reasonable usage of detection technology for tumor marker has direct effect on clinical tumor diagnosis and treatment. We were make a brief summary to actuality of the present applying tumor detection technologies in clinic and their developmental trends.

Objective To clone human survivin gene and express survivin-an inhibitor of apoptosis proteins in Pichia pastoris eukaryotic expression system. Methods Full length survivin gene was amplified by PCR using survivin specific primers. The verified survivin gene by sequencing was subcloned into pPic9k. The recombinant plasmid was linearized by restriction enzyme cutting. The linear survivin gene was introduced into GS115/ His-cells by electroporation. The transformants were transferred onto YPD plates that contained different concentrations of G418 for screening the positive clones. The integrated survivin gene in positive clones was confirmed by PCR. Selected transformants were cultured in BMMY medium with 1 % methanol for inducing the expression of survivin protein. The expressed survivin protein was analyzed by ELISA. Results The cloned human survivin sequence was the same as that in the GeneBank. The recombinant pPic9k-survivin was constructed in Picha pastoris eukaryotic expression vector. After the linear digestion, the recombinant vector was introduced into GS115/ His-cells, the 6 positive clones against G418(4 mg/mL) were obtained and confirmed by PCR; the highest survivin protein was expressed when expressed for two days in 1 % methanol BMMY medium. Conclusions Improved survivin protein yield may be reached by modifying the experimental conditions. This will help to further study the biological functions of survivin and its roles in tumor developments.

Objective: To investigate the possible association between the allelic variation of LAPTM4B and the genetic susceptibility of lung cancer. Methods: The genotype of LAPTM4B was analyzed in 134 unrelated healthy adult individuals and 166 patients with lung cancer by utilizing polymerase chain reaction based on special primers. The genotypical distribution of LAPTM4B was analyzed by ? 2 test. Results: The allelic frequencies of the *2 were 40.1% and 28.0% in the lung cancer group and the healthy control group respectively, which was significantly different between the two groups (P 0.002). There was a significant difference in the overall genotypical distribution between the patients and the controls (P0.005). The risk of suffering from lung cancer was increased 1.91 times in the individuals of the *1/2 genotype (95%CI: 1.178-3.110) and 3.26 times in the individuals of the *2/2 genotype of LAPTM4B (95%CI:1.338-7.929) compared with the *1/2 genotype. No association was observed between the genotypical distribution of LAPTM4B and the clinical information on patients of lung cancer such as gender, age, pathological type, differentiation classification of TNM and infection of HBV. Conclusion: This study suggests that the allele *2 of LAPTM4B might be the risk factor of lung cancer, which could be associated with genetic susceptibility of lung cancer.

Proliferative vitreoretinopathy (PVR)is a serious causing-blindness disease.It is intimately associated with retinal detachment (RD),proliferative diabetic retinopathy (PDR)and penetrating ocular injury with the complicated course and aggravating outcome.Proteomics is a science about investigating the total proteins in cellular level or organism level.The study of proteomics is helpful for reinforcing the cognition and understanding of pathogenesis and development of PVR.So proteomics study about PVR is becoming a new topic.The study progress in the mechanism of PVR,the study background and technique of proteomics,the application of proteomics study in PVR and relative ocular diseases mentioned above are reviewed in this paper.

Objective To study pathologic changes of gastric mucosa in duodenal ulcer(DU).Methods Pathologic changes of 650 cases of gastric mucosa in DU confirmed by pathology after subtotal gastrectomy were analyzed.Results There were 100% patients with complication of gastritis.Among them,506% patients accompanied with gastric mucosa chronic atrophic gastritis and 209% patients with gastric mucosa intestinal metaplasia.Conclusion DU is always accompanied by antral gastritis,the pathologic changes of gastric mucosa in DU relateed tightly with helicobacter pylori(Hp) infection and patient's age.

OBJECTIVE:To observe the clinical efficacy and transdermal absorption of Suhong bone-penetrating massage drug for rheumatic arthralgia. METHODS:Suhong bone-penetrating massage drug(Chinese medicine) was applied in 300 cases with rheumatic arthralgia characterized by various symptoms such as bone spur, injury, scapulohumeral periarthritis, acute injury or chronic strain of articular surrounding soft tissue; Suhongtougu powder was used as control to treat another 120 cases with the same disease. The curative efficacies in two groups were compared. The effects of the effective components such as Muskone,Borneo Camphor,Menthol in Suhong bone-penetrating massage drug on the transdermal absorption of this preparation was investigated by transdermal test on mice. RESULTS:The total effective rate in the treatment group was 93.24% as compared with 80.00% in the control group, showing significant difference between two groups(P

This study was aimed to investigate the effects of ursolic acid on human hepatic stellate cells (HSC-LX-2) proliferation and its mechanism.Different doses of ursolic acid were incubated with HSC-LX-2 cellin vitrof or 48 h.MTT was used for the detection of HSC-LX-2 cell proliferation.The expressions of PDGF-ERK signaling pathway associated proteins were measured by western blot.The results showed that the proliferation of HSC- LX-2 cells was inhibited by ursolic acid in a dose-dependent manner.The inhibition rate of 20,30 and 40μmol·L-1 of ursolic acid was 9.1%,42.3% and 62.6%,respectively.The IC50 was 35.2μmol·L-1.After incubated with ursolic acid for 48 h,protein levels of PDGF-R and p-ERK in 30 and 40μmol·L-1 group were significantly decreased when compared with the normal group (P<0.05 orP<0.01),except the ERK protein.It was concluded that ursolic acid can inhibit HSC-LX-2 cell proliferation.Its mechanism may be related to the blockage of PDGF-ERK signaling pathway.

Objective To establish a SYBR green Ⅰ real-time PCR method for detecting serum miR-21 and preliminarily explore its value in diagnosing breast cancer.Methods Total RNA was extracted from serum by Trizol reagent.Then miR-16 ( internal reference gene for miR-21 ) and miR-21 were reverse transcribed into corresponding cDNA by stem-loop RT primers.Their cDNA were amplified and detected by using SYBR green Ⅰ real-time PCR.The accuracy of assay was analyzed by signal to noise ratio (SNR) ; the specificity of assay was evaluated by melting curve; the precision of assay was assessed by R2 of standard curve and the stability of assay was calculated by intra-assay and inter-assay variation.Furthermore,the level of miR-21 and miR-16 were detected by this method among the serum samples of 33 breast cancer patients,18 benign breast disease patients and 49 healthy individuals.And the sensitivity and specificity in breast cancer diagnosis were evaluated according to cut-off value which was defined by relative expressions of miR-21 between breast cancer patients and healthy population.Results Through optimization of temperature and time in the annealing and extension stage during PCR,SNR was ≥99.36％ ; peak of melting curve was single; R2 of standard curve was 0.994 8 and Coefficient of Variance (CV) of intra-assay ＜ 1.5％,CV of inter-assay ＜4％.They indicated that this method was accurate,specific,precise and stable.When miR-16 was taken as internal reference,the relative expressions of serum miR-21 detected by SYBR green Ⅰ realtime PCR among the serum samples of breast cancer patients,benign breast disease patients and healthy population were 20.83 ± 18.18,20.86 ± 10.11 and 9.33 ±4.44,which had statistical significance among of them (x2 =16.92,P ＜ 0.001 ).There was statistical significance in healthy people vs breast cancer patients ( Z =- 2.58,P ≤ 0.01 ) and healthy people vs benign breast disease patients ( Z =- 4.42,P ≤0.01 ),but was not between breast cancer patients and benign breast cancer patients (Z =-0.51,P =0.608).When the value of 18.32 for the relative expressions of miR-21 was defined as cut-off value,the sensitivity and specificity of this method to diagnose breast cancer were 51.5％ (17/33)and 93.9％ (46/49),respectively.Conclusions A sensitive,specific and stable SYBR green Ⅰ real-time PCR for detecting serum miR-21 has been established.This method may have some diagnostic value for breast cancer.

Object To establish a HPLC method for determination of ethyl p hydroxybenzoate in Dragon's Blood. Methods Using C 18 reverse phase column and acetonitrile 1% acetic acid (31∶69) as mobile phase, detecting at 257 nm and quantitating with external standard method. Results The standard curves of ethyl p hydroxybenzoate showed good linearity over the range of 0.206-2.884 ng, r=0.999 8. The average recovery was 96.02% and RSD was 1.87%. Conclusion The method is quick and simple, and it is accurate and reliable for the determination of ethyl p hydroxybenzoate in Dragon's Blood.