Objective To evaluate the clinical significance of epidermal growth factor receptor EGFR) mutations in the treatment of non-small cell lung cancer ( NSCLC) . Methods Plasma DNAs solated fromblood specimens of 170 NSCLC patients, who were admitted in the First Affiliated Hospital of uangzhou Medical College from December 2005 to December 2007, were subjected to the test of EGFR utant-enriched PCR. The correlation of mutant detection with clinical characteristics was analyzed as well.Results Out of the total 170 patients, EGFR mutations were identified in 77 cases ( 77 /170, 45. 3% ) .EGFR mutations were more frequent in the patients with adenocarcinoma ( P lt; 0. 001) and in the nonsmokers P =0. 001) . In the 33 patients treated with gefitinib, those with mutations ( + ) showed a higher esponse rate and prolonged progression-free survival after the treatment compared with those with mutations( - ) ( P =0. 001 and 0. 001, respectively) . Conclusions EGFR active mutations can be specifically and ensitively detected by EGFR mutant enriched PCR assay. Plasma EGFR mutants detection is valuable in uiding clinical decision.
Objective To study the effect of chitosan (CS) mediated insul in-l ike growth factor 1 gene (igf-1) transfection on the repair of articular cartilage defect. Methods Twelve 3-month-old healthy male rabbits weighting 2.0-2.5 kg were randomly divided into 2 primary groups, control and intervention groups (n=6 per group). Control group was further divided into normal control (left knee) and normal saline (NS) control (right knee) groups. While, intervention group was divided into CS (left knee) and CS/igf-1 intervention (right knee) groups. Cartilage defects were created in the knee joints except normalcontrol. Intra-articular injections of CS/igf-1 complex was administrated 2 times a week for 4 weeks in CS/igf-1 interventiongroup, 0.5 mL CS in CS intervention group, and 0.5 mL sal ine solution in normal control and sal ine control groups. At 28days after treatments, the cartilage samples were collected for histological observation and collagen type II and aggrecan mRNA evaluation. Results HE staining and toluidine blue staining revealed that CS/igf-1 and CS intervention could significantly stimulated cartilage regeneration accompanied with fibrosis and inflammatory cell infiltration, however, CS/igf-1 treatment resulted in the best repair of cartilage defect. In contrast, sal ine control group only showed fibrous tissue prol iferation and inflammatory cell infiltration without significant cartilage repairing. In terms of collagen type II and aggrecan gene expression, significant differences were observed in each pairwised comparison among 4 groups in the order of CS/igf-1 gt; CS gt; NS gt; normal control (P lt; 0.05). Conclusion In situ CS/ifg-1 complex transfection can enhance the formation of mesochondrium by upregulating collagen type II or aggrecan expression, which might enhance the repair of articular cartilage defect.
Objective To investigate the gene expression of beta-defensin-4 (mBD-4) and mBD-6 in acute lung injury (ALI) mouse.Methods Sixty adult mice were randomly divided into a control group and a ALI group.ALI was induced by intraperitoneal injection of lipopolysaccharide (LPS) in the ALI group.The control group was treated with same dose of normal saline.The lung tissues were harvested at different time point after stimulation.The expression of mBD-4 and mBD-6 mRNA was measured by real-time quantitative reverse transcription polymerase chain reaction.DNA sequencing was used to confirm the specificity of mBD-4 and mBD-6 cDNA fragment.Results There were no obvious mBD-4 and mBD-6 mRNA expression in mouse lung in the control group at all time points and ALI 6 h group.In the ALI group a marked increasing expression was found on 12 h，1 d and 3 d after LPS stimulation.The mBD-4 mRNA expression was significant higher in the ALI groups of 1 d and 3 d points than that of ALI 12 h group with no obvious difference between each other.There were no significant differences of mBD-6 mRNA expression between ALI groups of 12 h，1 d and 3 d points Conclusion mBD-4 and mBD-6 mRNA is not constitutive expressed in mouse lung and show a up-regulative expression pattern after ALI.
ObjectiveTo detect expressions of PTEN and Ki-67 in primary thyroid cancer tissues and explore its clinical significances. MethodsThe expressions of PTEN protein and Ki-67 protein in 40 cases of paraffin-embedded tissues of primary thyroid cancer and the corresponding paracancerous tissues were detected by immunohistochemical method. The expressions of PTEN mRNA and Ki-67 mRNA in 14 cases of resected fresh tissues of primary thyroid cancer and the corresponding paracancerous tissues were detected by RT-PCR method. The relations between clinicopathologic characteristics and expression of PTEN protein or Ki-67 protein in the primary thyroid cancer tissues were analyzed. Results① The PTEN protein positive expression rate and the PTEN mRNA in the primary thyroid cancer tissues were significantly lower than those in the corresponding paracancerous tissues[35.0% (14/40) versus 60.0% (24/40), P<0.05; 0.225 7±0.036 3 versus 0.503 6±0.037 5, P<0.05], the Ki-67 protein positive expression rate and Ki-67 mRNA in the primary thyroid cancer tissues were significantly higher than those in the corresponding paracancerous tissues [72.5% (29/40) versus 42.5% (17/40), P<0.05; 1.212 1±0.042 1 versus 0.293 6±0.027 4, P<0.05]. ② The expressions of PTEN protein and Ki-67 protein were associated with the histological grading, pathological type, tumor stage, and presence of regional lymph node metastasis (P<0.05), which not associated with the patient's gender, age and integrity of tumor capsule or not (P>0.05). ③ The PTEN and Ki-67 protein expressions in the primary thyroid cancer tissues had a significantly negative correlation (rs=-0.605, P=0.000), which in the corresponding paracancerous tissues had no correlation (rs=-0.021, P=0.899). ConclusionPTEN and Ki-67 genes abnormally express in thyroid cancer tissue, which might be related with occurrence and development and its mechanism of primary thyroid cancer. Combination of two genes might contribute to identification of pathologic type, judge of biological behavior, and tumor stage of primary thyroid cancer, which might serve as a new target for diagnosis and treatment of it.
Objective To detect expression of Ras association domain family 1A （RASSF1A） gene in the colonic carcinoma tissue and to analyze the relationship of this expression to its clinical features. Methods Immunohistochemistry and Western blot methods were employed for detecting the RASSF1A protein expressions in 34 colonic carcinoma tissues and corresponding normal colon tissues. RT-PCR was employed for detecting RASSF1A mRNA expression. Results ①The RASSF1A protein expression in the colonic carcinoma tissues was significantly lower than that in the normal colontissues by using immunohistochemistry〔35.3% （12/34） versus 97.1% （33/34）， P＜0.05〕.There were significant relati-onships of RASSF1A protein expressions to the tumor differentiation and TNM stage （P＜0.05）， in other words， the positive rates of RASSF1A protein in the moderately and well differentiated andⅠ＋Ⅱof TNM colonic carcinoma tissues were all higher （P＜0.05）. ② The RASSF1A protein expression in the colonic carcinoma tissues was significantly lower than that in the normal colon tissues by using Western blot 〔0.316 8±0.019 6 versus 0.914 4±0.177 6， P＜0.05〕， which was close to the result of RT-PCR〔0.158 9±0.223 7 versus 0.572 3±0.193 9， P＜0.05〕. Conclusions Absentexpre-ssion of RASSF1A gene in the colonic carcinoma tissue might play an important role in tumor genesis and tumor progre-ssion， and it might become useful early detection of the colonic carcinoma.
Objective To explore the relationship between the HBsAg positive patients suffering from hepatocellular carcinoma (HCC) and HBV DNA genotype. Methods By using PCR type-specific primers combined with sequencing of genotype, we analyzed the genotype of HBV DNA in the serum of 500 patients with positive HBsAg in our hospital. Among them, 150 cases suffered from HCC. Results Genotype B and C were both predominant genotypes in HBsAg positive patients. But in HCC group, the rate of genotype C was 65.33% （98/150）, which was significantly higher than that in non-HCC group (88/350, 25.14%), while genotype B, in contrast, was 28.67% （43/150） and 68.86% （241/350）， χ2=75.45， Plt;0.05. The distribution of HBV DNA genotype B or genotype C in different gender or different age groups were not statistically significantly different in cases of HCC (Pgt;0.05). Conclusion Genotype C of HBV DNA is more common in patients with HCC, and maybe there is relationship between genotype C and the occurrence of HCC.
Objective To research the expressions of miR-196b and HoxB8 mRNA in colorectal cancer and theircorrelation with clinicopathologic features，and to explore the relationship between miR-196b and HoxB8 in vivo. Methods Expressions of RNA （including miR-196b and HoxB8 mRNA） and HoxB8 protein were detected respectively by using quantitative real-time reverse transcriptase PCR and Western blot in 30 cases of colorectal cancer and corresponding normalmucous membrane tissues. Results In colorectal cancer tissues，expressions of miR-196b and HoxB8 mRNA were higher than those of the corresponding normal mucous membrane tissues （P＜0.05）. Expression of miR196b mRNA was assoc-iated with lymph node metastasis，neoplasm stages （Ⅰ＋ⅡandⅢ＋Ⅳ），and distant metastasis （P＜0.05），on the otherhand，no significant differences were observed regarding tumor site，size，gross type，depth of invasion，tissue differentiation，age，and sex （P＞0.05）. Expression of HoxB8 mRNA was no significant differences concerning lymph node metastasis，tumor stages （Ⅰ＋Ⅱ，Ⅲ＋Ⅳ），distant metastasis，tumor site，size，gross type，depth of invasion，tissue differentiation，age，and sex （P＞0.05）. The expression of miR-196b mRNA was negatively correlated with HoxB8 mRNA expression （r=-0.458，P＜0.05），and HoxB8 protein expression with no obvious correlation （r=-0.236，P＞0.05） in colorectal cancer tissues. Conclusions The expressions of miR-196b and HoxB8 mRNA in colorectal cancer tissues are higher，the high expression of miR-196b mRNA is related to the tumorigenesis and progression of colorectal cancer as well as correlated with prognosis in colorectal cancer. The miR-196b inhibits the expression of HoxB8 mRNA by binding to the3′-UTR of target HoxB8 mRNA.
Objectives To analyze and assess the status of detection of human immune-deficiency virus (HIV) by PCR, and to find a new screening test of HIV. Methods Using the following keywords "diagnosis tests", "AIDS", "PCR" and "HIV", we searched the Medline and CBM from 1991 to 2001. Then we assess each of diagnosis test according to the international standards. Results 567 articles were searched, in which 53 articles were chosen to assess. In these 53 articles, it was found that 47% applied comparison with Golden Standard, 25% calculated sensitivity, 23% calculated specificity, and 23% calculated predictive value, no likelihood ratio was calculated in these articles. Conclusions It was still a kind of pilot-study to apply PCR to screening detection of HIV. The design methods of study should be improved.
One of the major clinical characteristics of congenital stationary night blindness(CSNB)is dysfunction of rod photoreceptors of the retina.Rhodopsin,the photosensitive pigment of the rods,is essential for maintaining the normal function of rod photoreceptors.It is resonable to hypothesize that mutations or deletions of rhodopsin gene may be involved in the molecular defect of CSNB.To test this hypothesis,we are searching for rhodopsin gene mutations in patients with autosomal dominant CSNB.In this study,DNA fragments containing the coding sequences in exon 5 of rhodopsin gene were amplified by polymerase chain reaction(PCR)in 15 patients and 5 unaffected members from a large family with autosomal dominant CSNB.RFLP analysis of these DNA fragments demonstrated that in comparison with a control group of 12 normal persons,there is no obvious deletion in exon 5 of rhodopsin gene,and that mutations or deletions do not exist in codon 314,codon 347,and the third base of codon 313 as well as the first base of codon 348 of the rhodopsin gene in these CSNB patients,which suggest the molecular pathogenesis of autosomal dominant CSNB not involve mutations or deletions of these codons of the rhodopsin gene. （Chin J Ocul Fundus Dis,1993,9:66-69）
ObjectiveTo detect the expression of Prox1 (prospero-related homeobox 1) gene in primary hepatocellular carcinoma (HCC), and to analyze the correlation of Prox1 gene expression with pathological grade and clinical stage of HCC. MethodsThe expressions of Prox1 gene in carcinoma tissues and adjacent cancerous tissues in HCC as well as normal liver tissues were detected by semi-quantitative RT-PCR, then the correlation of Prox1 gene expression with HCC pathological grade and clinical stage were analyzed. ResultsThe expression of Prox1 gene in carcinoma tissues (0.243±0.102) and adjacent cancerous liver tissues (0.537±0.235) was significantly lower than that in normal liver tissue (0.812±0.372), respectively （ Plt;0.01 or Plt;0.05）. Furthermore, the expression of Prox1 gene in carcinoma tissues was significantly lower than that adjacent cancerous liver tissues (Plt;0.05). The expressions of Prox1 gene in different pathological grade (F=97.950, Plt;0.001) and clinical stage were significantly different (F=228.300, Plt;0.001), and when compared with each other, the differences of pathological grade and clinical stage were also significant (Plt;0.001 or Plt;0.01). The expressions of Prox1 gene in HCC carcinoma tissue were negatively correlated with pathological grade (r=-0.930, Plt;0.01) and clinical stage (r=-0.980, Plt;0.01) of HCC. ConclusionsExpression of Prox1 gene may be related to the initiation and development of HCC, however, that whether Prox1 gene functions as tumor suppressor in HCC needs further investigation.