There are 3' non-coding region, downstream Rhesus box, SMP1 gene et cetera. after RHD stop code. This study was intended to determine the sequence of 3' non-coding region. One pairs of primer was designed and then a polymerase chain reaction (PCR) was established for specific amplification of whole length of 3' non-coding region of RHD in 10 Rh-positive and 10 D(el) samples. The PCR products were purified and directly sequenced. The results showed that all Rh-positive and D(el) samples were identical, which revealed that there were 103 bp between 3'-end of RHD coding region and 5'-end of downstream Rhesus box. The D(el) samples showed the same result with the normal Rh-positive sample. It suggests that lower expression of D antigen in D(el) red cells does not associate with 3' non-coding region of D(el) gene.
3' Untranslated Regions ; genetics ; Base Sequence ; Humans ; Molecular Sequence Data ; Rh-Hr Blood-Group System ; genetics ; Sequence Analysis, DNA

Objective To predict the CTL epitopes of Tat exon 1 region in HIV-1 CRF07_BC strains, which were prevailing in China. Methods Total of 236 plasma samples were from the 3rd National HIV Molecular Epidemic Survey (NMES3). All the subjects were infected with HIV-1 CRF07_BC viruses. The tat exon 1 region was amplified by reverse transcription reaction and nested polymerase chain reaction (nested-PCR), then the PCR products were sequenced. The distribution of CTL epitopes of this region were predicted by on-line software BIMAS HLA Peptide Binding Predictions and statistics software. Results To-tal of 236 CRF07_BC strains were from 16 provinces, mainly in intravenous drug asers(58.9%)and then sex(25.0%). It was showed that there were 12 CTL epitopes of 236 Tat exon 1 region of CRF07_BC strains mainly located in proline-rich region, cysteine-rich region and core-region. Those epitopes were banded by 5 HLA presenting molecules in genotype(A * 2501 ,A * 2902, B * 15,B * 5301 and Cw * 1203) and 6 HLA presenting molecules in serotype (B53, B58 ,B57 ,A3 ,A68 and Cw12). The frequency of single amino acid substitution was more than 50% in 7 CTL epitopes. Conclusion The CTL epitopes in Tat exon 1 of CRF07 _BC strains were located in different functional regions, and there were some amino acid variations in them.

Objective To verify the association between common breast cancer susceptibility loci which have been confirmed in European and Asian populations and breast cancer susceptibility in sporadic breast cancer among the Han nationality in Henan province , and to analyze their genotypes in the internal type of breast cancer . Methods In 253 breast cancer patients ( the case group ) and 343 patients who had benign breast lesions ( the control group), rs2046210(6q25.1), rs2981582(EGFR2), rs889312(MAP3K1), and rs3803662(TOX3/TNRC9)were genotyped by SNP im-LDR technique.According to estrogen receptor(ER), progesterone receptor (PR), human epidermal growth factor receptor 2(HER2)and Ki67, breast cancer are divided into 5 types:Lu-minal A, Luminal B, HER2-enrich, Luminal HER2, and triple negative breast cancer ( TNBC).Results rs2046210(6q25.1), rs2981582(EGFR2), rs889312(MAP3K1)had no statistical differences between the case group and the control group(P=0.421, 0.459, and 0.468), but the genotype of rs3803662(TOX3/TNRC9)in the case group and the control group had statistical difference (P=0.037).The allelic frequencies of rs3803662 between the case group and control group were different in codominant inheritance ( OR=2.19, 95%CI:1.19-4.02)and recessive genetic models (OR =2.06,95% CI:1.15 -3.70).Compared with AA and GA, GG in-creased the risk of breast cancer ( P =0.012, 0.015 ).The genotypes of rs2046210 ( 6q25.1 ), rs2981582 (EGFR2), rs889312(MAP3K1), and rs3803662(TOX3/TNRC9)had no difference in different types of breast cancer.Conclusions Four common breast cancer susceptibility loci from GWAS are not entirely associated with breast cancer risk among the Han nationality in Henan province .Only rs3803662(TOX3/TNRC9)is confirmed to increase the risk of breast cancer .Different genotypes of 4 loci distribute equally in different types of breast cancer .

Objective To study the expression and prognostic significance of neuroendocrine differentiation markers chromogranin A(CgA)and synaptophysin(SYN) in patients with resectable non-small cell lung cancer(NSCLC).Methods From January 2000 to January 2003,123 patients with NSCLC who received operations were investigated.The resected specimens and clinical data were collected.Immunohistochemical Elivison method was used to detecte the expression of CgA and SYN.Kaplan-Meier survival curve and Cox proportional hazard model multivariate analysis were applied for the prognostic factors. Results The positive expression rates of CgA and SYN were 22%,17.9%,respectively.The expression of SYN was associated with histological differentiation(P=0.001).No significant association was found between NSCLC with neuroendocrine differentiation(NSCLC-ND) and sex,age,smoke index,TNM Stage and pathology classification.No evidence showed the patients with positive expression of CgA or SYN could be tolerant with more cycles of chemotherapy(P=0.406).Kaplan-Meier survival curve indicated the survival had a relation with the expression of CgA and SYN.It was revealed by Cox analysis that SYN(P=0.001),TNM stage(P=0.02)and the maximal diameter of tumor(P=0.049) were independent prognostic factors. Conclusion The patients with NSCLC-ND had a poorer prognosis.SYN may be one of the prognostic factors in patients with NSCLC.

Objective To investigate the relationship between RAD51 135G ＞ C polymorphism and prognosis in triple-negative breast cancer patients by retrospective analysis.Methods The clinical data of 62 triplenegative breast cancer patients were collected.The 62 cases underwent standard chemotherapy and radiotherapy after tumor resection from Jan.2004 to Dec.2010 in Affiliated Cancer Hospital of Zhengzhou University.RAD51 135G ＞ C polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RELP) technology.The survival curve about progress free and overall survival time were then made.Results The median progress free and overall survival time in triple-negative breast cancer patients with or with-out RAD51 135G ＞ C polymorphism were(77.00 ±5.55)and(89.00 ± 10.40) months vs(99.00 ±4.26)and (103.00 ±4.30) months.The difference had statistical significance(P =0.039 and 0.015 respectively).Conclusion RAD51 135G ＞ C polymorphism is related with prognosis of triple-negative breast cancer patients,which might be a prognostic factor for breast cancer.

Objective To discuss the protein level of hypoxia-inducible factor 1-alpha(HIF-1α),glucose transporter-1 (Glut-1)and vascular endothelial growth factor(VEGF) in breast cancer tissue in diabetic patients and its significance.Methods HIF-1α,Glut-1 and VEGF protein levels were measured by immunohistochemical staining in 112 cases of primary breast cancer tissues.CD31 labeled vascular endothelial cells were used to evaluate micro vascular density (MVD).The correlation between the effect of blood sugar control and clinicopathological parameters was analyzed.Results The expression of HIF-1α,Glut-1 and VEGF protein in breast cancer tissues of diabetic patients was significantly higher than that in breast cancer tissues of non-diabetic patients(t =2.255,P =0.030; t =2.154,P=0.038; t =2.225,P =0.032).HIF-1α was positively correlated with Glut-1 and VEGF in diabetic patients with breast cancer (r =0.561,P =0.003 ; r =0.435,P =0.014).The level of MVD in breast cancer tissues of diabetic patients was obviously higher than that in the non-diabetic patients with breast cancer(t =9.458,P =0.000).The effect of blood sugar control was significantly correlated with lymph node metastasis and tumor stage in diabetic patients with breast cancer(x2 =4.689,P =0.030; x2 =5.051,P =0.025).Conclusion Hypoxia-related factors including HIF-1α,Glut-1 and VEGF and MVD are upregulated in diabetic patients with breast cancer,and the effect of blood sugar control is correlated with lymph node metastasis and tumor stage,suggesting diabetes mellitus may promote tumor progression through high glucose and hypoxia in breast cancer.

This review introduced the anti-tumor effects of non-steroid anti-inflammatory drugs (NSAIDs) and summarized their possible molecular mechanisms according to recent abroad literatures and our research results. Some evidence showed that the anti-tumor mechanisms of NSAIDs were different in various tumors.NSAIDs decreased the biosynthesis of PGE_2 and regulated the expressions of downstream correlated genes and proteins through restraining abnormal expression of COX-2 in certain neoplasms,which resulted in the inhibition of tumor angiogenesis and proliferation as well as induced apoptosis. But in other cancer cells, NSAIDs, as activators of peroxisome proliferator-activated receptor ? (PPAR?), induced COX-2 expression, promoted the biosynthesis of cyclopentenone prostaglandins (cyPGs). cyPGs further induced tumor cell apoptosis with PPAR? dependently or PPAR? independently. Since their special mechanisms of anti-proliferation and pro-apoptosis, NSAIDs revealed significant synergistic effects with other anti-tumor treatments.

Objective To compare three kinds of irradiation treatment plans for cervical and upper thoracic esophageal cancer,in order to arrived at proper decision for the patient.Methods From February 2001 to June 2004,43 such patients were studied with three different simulated treatment plans made including conformal plan,conventional four-field plan and conventional two-field plan for every one.All plans were evaluated with iso- dose curve and dose-volume histogram.Results GTV on 95% isodose curve was 99.5%,98.2% and 87.4% in conforaml plan,conventional four-field plan and conventional two-field plan,respectively;PTV_1 and PTV_2 on 95% isodose with 97.8%,97.2%,94.8% and 95.8%,86.6%,73.7%.The volume of＞20 Gy dose of left lung accepted was 18.6%,17.2% and 32.3%,in conformal plan,conventional four-field plan and conventional two-field plan,respectively;the right lung received 20.5%,19.9% and 35.5%.Conclusions Conformal plan is the best in radiotherapy,as it can provide ideal dose distribution of irradiated target with adequate protection of the normal tissues.Conventional four-field plan,being easy to carry out,can replace the conformal plan in most situations.Conventional two-field has the most uneven dose distribution and largest lung volume irradiated.

Highly Active Anti-Retroviral Therapy (HAART) has effectively inhibited the prevalence of HIV-1 and reduced the death rate caused by AIDS. In recent years,the emergence of resistance-conferring RT gene mutations in HIV-1 strains has become the major reason for HAART failure. The detection of drug resistance is important for the HAART regimen choice and novel drug development. A novel assay for the detection of HIV-1 RT drug resistance mutations was developed. HIV-1 drug resistance and wild strains in B subtypes were investigated using Two-Step Mutagenically-Separated PCR (MS-PCR),and point mutations including M41L,K70R,K103N,Y181C,T215F were detected. A longer mutant type primer was designed,using microplates hybridization and ELISA technique to detect several point mutations within a mixed mutant-wild type population. The results indicate that the Two-Step MS-PCR is as sensitive and specific as that in the traditional MS-PCR and MS-PCR combined with ELISA can give a good P/N quotient with better sensitivity,low cost,relatively less time consumption and high-throughput screening. It will be used in clinic usage for the detection of HIV-1 drug resistance mutations as well as other point mutations.

HIV-1 envelope glycoprotein gp41,which is a hopeful target for HIV-1 fusion inhibitors,plays a critical role in the fusion of viral and cellular membranes.In order to build up the screening assay of HIV-1 fusion inhibitors targeting gp41,HIV-1 gp41 5-helix and 6-helix were expressed in prokaryotic cells.Gp41 5-helix and 6-helix recombined plasmids were constructed by using PCR,enzyme digestion and ligation taking the clade B HIV-1 genome as a template.The plasmid was transferred into E.coli BL21(DE3)and then induced by IPTG.The expressed protein was purified by affinity chromatography after denaturation and renaturation.The SDS-PAGE analysis was used during expression and purification.Native-PAGE was used to identify the interaction between gp41 5-helix and T-20.The result will be helpful to build up the screening assay of HIV-1 fusion inhibitors targeting gp41.