Mutation of ras gene family was studied in 206 cases of gastric cancer and precancerous lesions by PCR/RFLP, PCR/SSCP and DNA suquencmg. The results showed: the mutation rates of H-ras 12 codon in metaplasia, atypical dysplasia, early cancer and advanced cancer were 16.7%(6/36), 31.2%(15/48), 50.0% (5/10), 32.2% (20/60), respectively. In the groups of superficial gastritis and normal controls, no mutations were found. Mutations of H-ras 61 codon and N-ras 12 codon in various groups were the same as that of normal controls. Only 2 cases of K-ras 12 codon mutation were detected in gastric cancer group by PCR-SSCP, but it was not identified by DNA sequencing. It might be due to polymorphism. All H-ras 12 codon mutations were of G→T mutation. There were significant differences between groups of metaplasia, dysplasia, gastric carcinoma and normal controls. (P

Objective:This study was aimed to investigate the influence of CDK7 siRNA on the sensitivity of endometrial carci-noma cell line HEC-1-A to cisplatin (DDP)-based chemotherapy. Methods:Different CDK7 siRNA fragments were synthesized based on the designs of the CDK gene sequence and were transfected into HEC-1-A. Real time reverse transcription polymerase chain reac-tion (RT-PCR) and Western blot analysis were employed to demonstrate the effects of transfection. The best CDK7 siRNA was chosen to specifically silence CDK7 expression in HEC-1-A.The sensitivity of the cells to DDP therapy before and after transfection was deter-mined by methyl thiazol tetrazolium (MTT) cytotoxicity assay, flow cytometry, and Hoechst/PI double-staining fluorescence microsco-py. Results:A total of four different CDK7 siRNA segments were designed and successfully transfected into HEC-1-A cells. The inter-ference effect in each group was confirmed by real time RT-PCR and Western blot assays. CDK7-423 was determined as the best per-forming CDK7 siRNA (over 70%) to transfect into HEC-1-A cells. MTT cytotoxicity test showed that IC50 of DDP decreased to a range from 45.122 μg/mL and 3.200 μg/mL after inhibition of CDK7 expression. DDP toxicity to the endometrial carcinoma cells sig-nificantly increased (P<0.05). Flow cytometry revealed that the average cell apoptosis rate significantly increased after the inhibition of CDK7 expression (11.66%to 37.57%, P<0.05). Similar results were observed using Hoechst/PI double-staining fluorescence microsco-py, and the number of apoptotic corpuscle demonstrated an apparent increase in the low CDK7-expressing group compared with the pa-rental cells. Conclusion:After the downregulation of CDK7 expression by CDK7 siRNA transfection, DDP chemotherapy sensitivity and apoptosis of endometrial carcinoma cells significantly increased. Further research is anticipated on the use of CDK7 as a new treat-ment target for endometrial carcinoma.

Objective To explore the effect and molecular mechanism of N-myc downstream regulated gene 1 (NDRG1) on transforming growth factor-beta (TGF-β)-induced epithelial-mesenchymal transition (EMT) in human lung cancer cells.Methods Lung cancer A549 cells were transfected with NDRG1 overexpressed vector,and then treated with 5 μg/L TGF-β1.The abilities of invasion were detected by Transwell assay.The expressions of NDRG1 mRNA and protein were analyzed by teal-time reverse transcription polymerase chain reaction (RT-PCR) were examined with Western blot.The expressions of EMT-associated markers E-cadherin and Vimentin,and EMT-associated signaling molecules Snail,AKT and Smad were detected with Western blot.Results We found that TGF-β1 treatment could induce morphological alteration of A549 cells from epithelial morphology to mesenchymal morphology.TGF-β1 significantly increased the migration of A549 cells,and increased the expression of mesenchymal maker vimentin and decreased epithelial marker E-cadherin.More importantly,overexpression of NDRG1 significandy reversed the effects of TGF-β1 on A549 cells.Moreover,NDRG1 significandy decreased the levels of phospho-AKT,and suppressed the expression of EMT-related transcription factor Snail.Conclusions NDRG1 could reverse the effects of TGF-β1 on EMT in A549 cells,by which mechanism is related to reduction of the expressions of phospho-AKT and Snail.

Objective To observe the protection of Caspase inhibitor(zVAD-fmk,benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone) on neonatal rat with hypoxic-ischemic brain damage(HIBD).Methods Thirty-six neonatal rats,7 days old,were randomly divided into hypoxic-ischemic(HI) control group(A),zVAD-fmk treated group(B) and sham group(C).Before HI insult,a pan-Caspase inhibitor,zVAD-fmk or normal buffer solution was injected into the cerebral ventricle.The water content of cerebral hemisphere was measured and the percentage of apoptofic cells in hippocampal neurons was measured by Flow cytometer(Annexin V/PI) at 24 hours after HI insult.The effect on body weights(percentage of increased weight,WIP) and macroscopical changes(percentage of cortox and hippocampal dead neurons) were assessed at 14 days.Results Compared with group A,the water content of ischemic hemisphere and apoptosis percentage of hippocampal neurons in group B reduced significantly.The difference of percentage of increased weight at 14 days in group B was not significantly.Microscopic examination showed that cortox and hippocampus neural death rate in group B was proved significantly reduced compared with that in group A.Conclusion Intracerebral administration of zVAD-fmk has protective effects on hypoxic-ischemic brain damage in neonatal rat.

Glycyrrhiza uralensis Fisch. ex DC is widely used in traditional Chinese medicine (TCM). Among its various active components, glycyrrhizic acid is believed to be the marker component. Squalene synthase (SQS) and beta-amyrin synthase (beta-AS) are key enzymes in the biosynthetic pathway of glycyrrhizic acid in G uralensis. To reveal the effects of co-expression of SQS1 and beta-AS genes on this pathway, 7 yeast expression vectors harboring different SQS1 variants and beta-AS were constructed and expressed in Saccharomyces cerevisiae as fusion proteins. TLC and GC-MS results showed that co-expression of SQS1 and beta-AS enhanced the accumulation of beta-amyrin. The effects of SQS12 were more obvious than the other two SQS1 variants. This study is significant for further investigations concerned with exploring the biosynthesis of glycyrrhizic acid in vitro and strengthening the efficacy of G. uralensis by means of increasing the content of glycyrrhizic acid.
Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Glycyrrhiza uralensis ; genetics ; Intramolecular Transferases ; metabolism ; Oleanolic Acid ; analogs & derivatives ; metabolism ; Plant Proteins ; genetics ; Recombinant Proteins ; metabolism ; Saccharomyces cerevisiae ; metabolism

Objective To assess the anatomic morphology of the connective structures among brain ventricles and cerebrospinal fluid (CSF) movement in them by CSF spin-labeling MR imaging. Methods According to the order of registration, 50 healthy volunteers were randomly selected and received cerebrospinal fluid spin-labeling MR scan with time-spatial labeling inversion recovery single-shot fast spin echo sequence (SLIR-SSFSE). The tagged CSF was used as an endogenous tracer. The anatomic morphology of the connective structures of brain ventricles and the flow direction of CSF were observed. The longitudinal diameter and transverse diameter of bilateral foramina of monro, midbrain aqueduct, and the central and bilateral lateral apertures of the fourth ventricle of each subject were measured and calculated based on multiple measurements. The flow rate of CSF was calculated based on the flow distance of CSF in the connective structures between brain ventricles during different TI time. The mean value of each indicator was acquired. Results Two-way flow state of CSF was observed in all connective structures, including bilateral foramina of monro, midbrain aqueduct, and the central and bilateral lateral apertures of the fourth ventricle. On the coronal planes, foramen of monro appears as a "Y"-type tubular structure locating among the both sides of the anteriomedial thalamus and fornix, which connect upward with bilateral lateral ventricles and downward with the third ventricle. The longitudinal diameter and transverse diameter of the left side of foramen of monro were 3.50-5.50 mm[mean (4.37 ±0.47)mm]and 1.00-1.40 mm[mean(1.21 ±0. 13) mm], respectively. The longitudinal diameter and transverse diameter of the right side of foramen of monro were 4. 20-4. 80 mm[mean(4.42 ± 0.20) mm]and 1.00-1.60 mm[mean (1.21 ±0. 19) mm], respectively. On the sagittal planes, foramen of monro appeared as an oblique fine tubular structure with the angle of 55°-58° between the both sides. CSF flow velocity towards the foot was 1.61-2. 52 mm/s[mean (2. 00 ± 0. 17) mm/s]in the left side of foramen of monro and 1.93-2. 20 mm/s [mean (2.03 ±0.09) mm/s]in the right side of foramen of monro. On the sagittal planes, midbrain aqueduct appeared as a curved tubular structure through diencephalon, with the longitudinal diameter of 9.90-17.30 mm[mean(15.51 ± 1.70) mm]and the transverse diameter of 1.70-2.30 mm[mean (1.92 ± 0. 17)mm]. In midbrain aqueduct, CSF flow velocity towards the foot was about 5.00-8.74 mm/s[mean (7.84 ±0.86) mm/s]and towards the head was about 3.84-6.71 mm/s[mean (6. 01 ±0. 66) mm/s]. On the sagittal plane, the central apertures of the fourth ventricle appeared as a fork-like tubular structure, with a small bifurcation forward to the central canal of the medulla oblongata and a posterior branch downward to cerebellomedullary cistern. On the coronal plane, lateral apertures of the fourth ventricle appeaed as curved tubular structures connecting the lateral horn of the fourth ventricle and cerebellomedullary cistern. The longitudinal diameter and transverse diameter of the left lateral aperture were 6.30-14. 60 mm[mean (10.42 ±2.88) mm]and 1.00-1.50 mm[mean (1.24 ±0.18) mm],respectively; of the right lateral aperture, they were 6. 20-15.50 mm[mean (12. 13 ± 3.05) mm]and 1.00-1.40 mm[mean(1.19 ±0. 13) mm], respectively. The angle range between the left and right lateral aperture was from 87° to 114°. CSF flow velocity towards the foot was about 2. 89-6. 70 mm/s[mean (4. 78 ± 1.32) mm/s]in the left lateral aperture and 2. 84-7.11 mm/s[mean (5.56 ±1.40) mm/s]in the right lateral aperture. Conclusions CSF spin-labeling MR imaging could display the anatomic morphology of the connective structure among brain ventricles and could be used for noninvasively assessing CSF movement.

Objective To investigate the surgical selection of retinal detachment with myopic macular holes. Methods Management strategies were adopted according to the ocular symptoms of patients with high myopia involved in posterior chorioretinal atrophy, the colour showed by the bottom of macular holes and the shape of posterior staphyloma. Seventy-five eyes of 72 patients fulfilled the study criteria. The patients were followed up in an average of eight-month-observation and were divided into three groups: single 16% C2F6 gas tamponade (26 eyes of 26 patients) as group A,vitrectomy combined with 16% C2F6 gas tamponade (26 eyes of 23 patients) as group B and vitrectomy with silicon oil tamponade (23 eyes of 23 patients) as group C. Results 69.2%(18/26) of the macular holes were closed in group A, 73.1%(19/26)in group B and 87.0% (20/23) in group C after the primary surgery. Comparison of postoperative outcomes,macular hole closure rate was not significantly different among three groups (P= 0.3184). Conclusions It suggests that good anatomic retinal reattachment be achieved using any approach. However, the optimal surgical choice regarding of the personality case offered less surgical burden and more rapid visual rehabilitation to patients.

Objective To explore the role of forkhead box protein O3a (FOXO3a) in inhibiting the proliferation and metastasis of breast cancer cells by downregulation of vascular endothelial growth factorA (VEGF-A).Methods Cell proliferation bioassay and plate clone formation assay were used to compare the changes of proliferation after siRNA interference.Wound healing and Transwell were used to compare the changes of invasion metastasis after interference.Meanwhile,the changes of VEGF-A expression in the cells before and after interference were compared by real time polymerase chain reaction (RT-PCR) and Western blot.Furthermore,the expressions of mRNA FOXO3a and VEGF-A were detected in 20 cases of breast cancer patients in breast cancer tissues and adjacent normal breast tissues.Results The MCF-7 and MDA-MB-231 cells were increased in cell proliferation and invasion after interference.Further studies found that mRNA and protein expression of VEGF-A were up-regulated in MCF-7 and MDA-MB-231cells after interference.In vivo the expression of FOXO3a was lower in 15 cases of cancer compared to normal tissues,and the expression of VEGF-A was high in 15 cases of cancer (75％).FOXO3a and VEGF-A expression was highly negatively correlated.Conclusions This study showed that FOXO3a could inhibit the proliferation and invasion of breast cancer cells,which might be regulated by VEGF-A.It provides an important theoretical evidence for targeted inhibition of breast cancer metastasis.

Female ; Humans ; Obesity ; complications ; Polycystic Ovary Syndrome ; complications ; therapy

Objective To evaluate the efficacy and toxicity of the docetaxel combined with nedaplatin or cisplatin in the treatment of patients with advanced head and neck carcinoma.Methods 58 patients with advanced head and neck carcinoma were enrolled into the study.These patients were divided into nedaplatin group (27 patients:docetaxel 75mg/m2 on day 1 and nedaplatin 80mg/m2 on day 1 of the 21-day cycle) and cisplantin group( 31 patients:docetaxel 75mg/m2 on day 1 and cisplatin 25mg/m2 on day 1-3 day of 21-day cycle).Each patient should complete two cycle.Results The response rate in nedaplatin group was 59.2％ and in cisplantin group was 54.8％,the difference was statistically significant between the two groups( P ＞ 0.05 ).The rate of white blood cell and plate let reduction in nedaplatin group was 55.5％,40.7％,respectively,and those in cisplantin group was 51.6％,35.4％,respectively,the difference was statistically significant between the two groups ( call P ＜ 0.05 ).The rate of vomit in nedaplatin group was 29.6％,which was lower than that of cisplantin group (64.5％) ( x2 =4.02,P ＜ 0.05 ).Conclusion In the treatment of advanced head and neck carcinoma,the combination of docetaxel and nedaplatin had the same efficacy as the combination of docetaxel and cisplatin,and they appeared to be more-tolerated.