Objective: To investigate the biological significance of the expressions of CDC25A and CDC25B and their relation to cell apoptosis and proliferation of esophageal squamous cell carcinoma. Methods: The expressions of CDC25A and CDC25B were examined by SP immunohistochemical stain (IHC) in 52 cases of esophageal carcinoma specimens, 25 cases of dysplasia and 24 cases of normal tissues. The apoptotic ratio, proliferation index (PI), the expression of CDC25A and CDC25B protein were determined by FCM in 30 cases with esophageal carcinoma and 5 cases of normal tissues. Results: The positive expression rates of CDC25A (50%) and CDC25B (48.1%) in esophageal carcinoma were significantly higher than those in dysplasia(16%) and normal tissues (0%) (P < 0.01); the expressions of CDC25A and CDC25B were correlated with differentiation degree and invasion depth of tumor cells, and CDC25A was related with lymph node metastasis of esophageal squamous cell carcinoma. The PI was significantly higher in nuclear membrane protein CDC25A-positive group than that in nuclear membrane protein CDC25A-negative group (P < 0.05). Conclusion: CDC25A is involved in the carcinogenesis and progression of esophageal squamous cell carcinoma and has the potential to become a new biological marker to predict the carcinogenesis and prognosis of esophageal carcinoma; CDC25B might play a role in the early phase of esophageal squamous cell carcinoma. The CDC25A protein in the nuclear membrane might contribute to cell proliferation.

OBJECTIVE: To construct a plant effective expression vector driven by a fruit specific promoter for the expression of hepatitis B virus surface antigen (HBsAg), to further improve the expression of exogenous gene in plant, and to prepare for the development of an effective anti-hepatitis vaccine. METHODS: Tomato fruit-specific promoters' gene 2A12 and E8 were respectively introduced to pBPFOmega7 to form pB2A12 and pBE8. The DNA fragment containing HBsAg-s gene from plasmid YEP-HBs was inserted respectively into pB2A12 and pBE8 to form pB2A12-HBs and pBE8-HBs. The fragment containing "p35S+2A12+Omega+HBsAg-s+Tnos" of the pB2A12-HBs was sub-cloned into plasmid pCAMBIA1301 to yield the reconstructed plant binary expression plasmid pCAM2A12-HBs, and the fragment containing "p35S+E8+Omega+HBsAg-s+Tnos" of the pBE8-HBs was sub-cloned into plasmid pCAMBIA1301 to yield the plasmid pCAME8-HBs. The inserted gene HBsAg and fruit-specific promoters in the reconstructed plant binary expression vectors were confirmed by sequencing. Then, pCAM2A12-HBs and pCAME8-HBs were directly introduced into Agrobacterium tumefaciens strain EHA105. RESULTS: Digestion with restriction enzymes proved that all recombinant vectors had the inserts with expected length of the target fragments, and the sequencing results were confirmed correct. CONCLUSION: In this study, plant expression vector containing HBsAg gene driven by fruit specific promoter and CaMV35s promoter was successfully constructed.

In recent years traditional Chinese medical massage has made remarkable achievements in the treatment of infantile diarrhea, besides, it has no toxic and side effect, no complaints and without affecting the growth rhythm of children and easily accepted by children. Massage manipulation and curative effects of treating infantile diarrhea was discussed in this text, the mechanism was analyzed, and the forecast of traditional Chinese medical massage on infantile diarrhea was proposed.

Objective To establish a HPLC method for determination of benzylacetone in the rhododendron.Methods The HPLC analysis was carried out on Luna C18(250 mm×4.6 mm,5 μm) at 30℃temperature.The mobile phase was acetonitrile-water (64 ∶ 36),the flow rate was 1 ml/min,and the detection wavelength was 220 nm.Results The linear range of benzylacetone was 17.08～170.8 μg/ml (R=0.9998),the average recovery of this method was 99.81％,and the RSD was 1.46％.Conclusion The method is convenient,quickly and accurate,can be used for the quality control of rhododendron oil and their preparations.

Objective To explore the relationship between methylenetetrahydrofolate reductase (MTHFR) 677C ＞ T and unexplained recurrent pregnancy loss (URPL).Methods All patients were recruited from the outpatient department of Obstetrics/Gynaecology & Genetics of Hangzhou First People's Hospital from January 2013 to May 2014.A case-control study was performed.According to the stochastic indicator method,there were 125 subjects with a history of ≥2 times URPL as the case group,and 905 healthy parous women with no history of URPL as the control group.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the distributions of the polymorphisms of MTHFR 677C ＞ T,and the results were validated using oligo-chip and direct sequencing.Results The allele and genotype frequencies of MTHFR were 60.0％ for C,40.0％ for T,32.0％ for CC,56.0％ for CT,and 12.0％ for TT in the case group and 67.4％ for C,32.6％ for T,41.4％ for CC,52.0％ for CT,and 6.6％ for TT in the control group,respectively.The prevalence of allele T was significantly higher in the case group than in the control group (OR =1.379,95％ CI =1.051-1.808,P =0.020),the frequency of genotype TT was also significantly higher in the case group than in the control group (OR =2.344,95％ CI =1.220-4.503,P =0.009).Conclusion The fertile women with MTHFR 677T allele and 677TT genotype may be susceptibility to URPL in a Chinese Han population from the Hangzhou area.

OBJECTIVETo study the effect of aqueous extract of Taxus chinensis var. mairei (AETC) combined Erlotnib on the growth of A549 xenograft in nude mice and its mechanism.

METHODSThe xenograft model in nude mice was established by inoculating A549 cells subcutaneously. BALB/c nude mice bearing A549 xenograft were randomly divided into six groups, i.e., the low dose Erlotinib group (A) , the standard dose Erlotnib group (B) , the low dose Erlotinib combined AETC group (C), the standard dose Erlotnib combined AETC group (D), the AETC group (E), the control group (F), 12 in each group. Different medication was performed for 7 successive weeks after 24 h. One mL blood was withdrawn and tumor tissues taken. The tumor inhibition rate was calculated. The combined effect was analyzed by Jin's Formula [Q = Ea + b/(Ea + Eb-Ea x Eb) ]. mRNA and protein expression levels of epidermal growth factor receptor (EGFR), cyclooxygenase-2 (COX-2), and B cell lymphoma-2 (Bcl-2) in xenografts were detected using real-time RT-PCR and ELISA.

RESULTSCompared with Group F, the xenograft weight was obviously lowered in Group B-E (P < 0.05, P < 0.01). The q value was 0.92 in Group C and 0.96 in Group D, which was obtained by simple adding of the two drugs. Compared with Group F, EG- FR mRNA expression in Group D and E, COX-2 mRNA expression in Group A-E; Bcl-2 mRNA expression in Group B-D; COX-2 protein expression in Group B-E; Bcl-2 protein expression in Group C and D were obviously lowered with statistical difference (P < 0.05, P < 0.01).

CONCLUSIONSAETC combined low dose and standard dose Erlotinib had synergistic effect on tumor inhibition. Its mechanism might be associated with down-regulating mRNA and protein expression levels of COX-2 and Bcl-2.

Animals ; Antineoplastic Agents ; pharmacology ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Cell Line, Tumor ; Cyclooxygenase 2 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Erlotinib Hydrochloride ; pharmacology ; Heterografts ; Lung Neoplasms ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Receptor, Epidermal Growth Factor ; metabolism ; Taxus ; Transplantation, Heterologous

Adult ; Case-Control Studies ; Comet Assay ; DNA Mutational Analysis ; Female ; Humans ; Male ; Micronucleus Tests ; Middle Aged ; Occupational Exposure ; Vincristine

Animals ; Brain ; metabolism ; Female ; Ginkgolides ; pharmacology ; Hypoxia ; metabolism ; Lactic Acid ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; metabolism

Branchio-Oto-Renal Syndrome ; Child ; Humans ; Male

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Liver Cirrhosis ; congenital ; pathology ; Male