【Abstract】Objective To investigate the expression of the mRNA of cancer-testis antigen 9 (CT9) gene in hepatocellular carcinoma. Methods The expression of CT9 mRNA was detected through RT-PCR in HCC tissues and their adjacent non-HCC tissues from 45 HCC patients. From CT9 RT-PCR positive products, 3 samples were selected randomly and were sequenced. ResultsCT9 mRNA was detectable in 51.1%(23/45) of HCC samples, and no expression of CT9 mRNA was detected in the adjacent non-HCC tissues. In addition, the RTPCR products were proved to be CT9 cDNA by DNA sequencing. No relationship was found between the expression of CT9 mRNA and clinical factors such as age, sex, tumor size, degree of tumor differentiation, serum αfetoprotein level and infection of hepatitis B virus or hepatitis C virus （Pgt;0.05）. ConclusionCT9 mRNA is expressed with high percentage and specificity in hepatocellular carcinomas. The CT9 gene product is a potential target for antigenspecific immunotherapy of HCC.
Objective To evaluate branched-chain DNA (b-DNA) signal amplification and semi-quantitative (Sq) RT-PCR in detection of free cancer cells in peritoneal flushing fluid of colorectal cancer patients during surgery. Methods The CEA mRNA in peritoneal flushing fluid in 48 cases of colorectal cancer were detected by b-DNA and SqRT-PCR. Peritoneal flushing fluid cytology (PLC) was conformed simultaneously to detect the free cancer cells. The peritoneal flushing fluid of 12 cases with colorectal benign disease were taken as negative control, GAPDH mRNA as internal control. Results In colorectal cancer patients, positive rate of free cancer cells by bDNA and SqRT-PCR （43.8%, 31.3%） was higher than that by PLC （4.2%）. The relative quantitative expressions of CEA mRNA were related to the Dukes staging， depth invasion and differentiation degree (Plt;0.05)， but irrelevant to tumor size，the patients’ age and gender (Pgt;0.05).Conclusion Both b-DNA and SqRT-PCR technologies have advantages and disadvantages to detect free cancer cells in peritoneal flushing fluid， which are related to clinicopathological factors.
Objective To observe the effect of gefitinib on expression of epidermal growth factor receptor (EGFR) in bile duct epithelial cells, and the feasibility of inhibiting hyperplasia of bile duct epithelial cells with gefitinib. Methods Sixty-one patients with hepatolithiasis having to be in hospital for surgery from the First People’s Hospital of Shuangliu county were selected, with 25-65 years old, average 46.92 years. The patients were randomly divided into therapy group and control group. There were 30 cases in therapy group, in which fine duct was placed on lesion bile duct during operation, and through whom gefitinib solution was perfused after operation. There were 31 cases in control group with only T tube drainage after operation. The bile duct sample was obtained respectively during the operation and 6 weeks and 12 weeks after operation. The histology and expression change of EGFR were observed by HE staining, immunohistochemistry and RT-PCR method respectively. Results There were no significant differences in pathohistology changes of bile duct and the EGFR protein and mRNA expression between therapy group and control group during operation. The hyperplasia of epithelium mucosae and submucosal gland in the therapy group were obviously decreased as compared with those in control group, the EGFR mRNA and protein expression in therapy group were weaker than those of control group （Plt;0.05） 6 weeks and 12 weeks after gefitinib treatment. Conclusion EGFR is overexpressed in the chronic proliferative cholangitis, and continuously local application of gefitinib after operation can specifically interrupt the activation and expression of EFGR and then effectively inhibit the hyperplasia of bile duct epithelial cells.
Objective To evaluate the expressive varieties of Nogo-A mRNA in injured optic nerves of rats. Methods Reverse transcription polymerase chain reaction (RT-PCR) method was used to hemi-quantitatively analyze the levels of Nogo-A mRNA in the optic nerves 3, 7, 9, 15, 21, and 25 days respectively after injury.Results The level of the expression of Nogo-A mRNA was low in the normal optic nerves, while it was significantly high in the optic nerves 3 days after in jury, and kept the high level still after 25 days.Conclusion The expression of Nogo-A mRNA in injured optic nerves is increased. (Chin J Ocul Fundus Dis，2003，19：201-268)
ObjectiveTo detect the 2019 novel coronavirus (2019-nCoV) in various biological specimens of novel coronavirus pneumonia (NCP), and preliminarily observe the status of 2019-nCoV in different systems of the body and its clinical significance.MethodsThe study design was a small-scale cross-sectional observational study. All the confirmed NCP cases being treated in the Second People’s Hospital of Yibin · West China Yibin Hospital, Sichuan University on February 2nd, 2020 were enrolled in this study. Two sets of primers were designed for 2019-nCoV-1ab and N regions using real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay. The 2019-nCoV in upper respiratory specimens, blood, feces and urine specimens of the NCP cases were detected on the single day.ResultsA total of 7 imported NCP cases (mild type) were included. The 7 patients were confirmed by the positive results of 2019-nCoV nucleic acid tests of upper or lower respiratory specimens between the 3rd day and the 7th day after fever onset, while 2 patients were found positive on the 3rd day after onset. The 2019-nCoV nucleic acid tests of the 7 patients were detected again on a single day between the 7th day and the 15th day after onset, and the results showed: the upper respiratory specimens of 5 patients were found negative (1 case was on the 7th day after onset); 2019-nCoV was not detected in the blood, feces or urine specimens of the 7 patients.ConclusionsFor mild type NCP patients, real-time RT-PCR test could detect 2019-nCoV between the 3rd day and the 7th day after onset, while 2019-nCoV might become negative since the 7th day after onset. 2019-nCoV was not detected in the blood, feces or urine of mild type NCP patients on the single day between the 7th day and the 15th day after onset. This study was only a preliminary observational study, which needed high-qualified studies to obtain more definitive conclusions.
【Abstract】 Objective To investigate the expression and significance of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors in pancreatic cancer. Methods Thirty-two samples of pancreatic cancer tissue were collected from year 2002 to 2004. All of them were verified by histopathology and there were 9 cases of well-differentiated, 12 of moderately differentiated, and 11 of poorly differentiated, in which 12 cases were in the stage of Ⅰor Ⅱand 20 in the stage of Ⅲ or Ⅳ according to the TNM staging method. Eighteen normal pancreatic tissues were used as control group. The expressions of TRAIL receptors (death receptor 4, death receptor 5, decoy receptor 4 and decoy receptor 5) mRNA were assayed by semi-quantitive reverse transcription polymerase chain reaction (RT-PCR) in the pancreatic cancer tissues and the normal pancreatic tissues. Results The expressions of death receptor 4 (DR4) and death receptor 5 (DR5) were detected in all the pancreatic cancer tissues and the normal pancreatic tissues and the levels of DR4 and DR5 were significantly higher than those of the normal pancreatic tissues (P＜0.01). Decoy receptor 1 (DcR1) and decoy receptor 2 (DcR2) were also expressed in normal pancreatic tissues, whereas DcR1 and DcR2 were only expressed in 18 and 20 pancreatic cancer tissues, respectively. However, there were no significant difference of the expression of DcR1 and DcR2 between the pancreatic cancer tissues and the normal pancreatic tissues （Pgt;0.05）. The expression level of DR5 in pancreatic cancer tissue was correlated with tumor differentiation and clinical stage, and the levels in stage Ⅲ and stage Ⅳwere significantly lower than those of stageⅠand stageⅡ(P＜0.05). The expressions of DR4, DcR1 and DcR2 were not correlated with tumor differentiation and clinical stage (Pgt;0.05). Conclusion ①The expression of TRAIL receptors in pancreatic cancer tissues is prevalent, but the types of receptors expressed in different tissues were also different. High expression of death receptors may play an important role in TRAIL recptors regulated pancreatic cancer apoptosis. ②The expression of DR5 is correlated with the differentiation degree of pancreatic cancer cell and clinical stage of tumor. The expressions of DR4, DcR1 and DcR2 should not be considered as related indexes of differentiation degree or clinical stage of pancreatic cancer.
Objective To verify whetheriris pigment epithelial cells(IPECs)possess the similar potential of specific phagocytosis to retinal outer segments(ROS) with retinal pigment epithelialcells(RPECs). Methods IPESc were isolated from neonatal bovines with Hu's method,and were cultured.The cultured cells were identified by immunohistochemical methods with antibodies to cytokeratin and s-100.Total RNA of IPECs was extracted by Trizol.The specific primers for mannose-receptor andbeta;-actin were designed according to their sequence from Genbank.The mRNA expression of these proteins in the IPECs was analyzed by reverse transcription polymerase chain-reaction (RT-PCT).Results The Cultured IPECs have no contamination of other cells .The extracted RNA was ideal and had no degradation.RT-PCR analysis showed that mannose-receptor's mRNA was expressed in cultured IPECs in vitro.ConclusionCultured IPECs may express the mannose-receptor,and may have similar potential of phagocytosis to ROS with REPCs.
【Abstract】ObjectiveTo discuss the possibility and clinical significance of SSX-1 mRNA used as specific marker for examining HCC cells in peripheral blood of HCC patients. MethodsUsing the RT-PCR method, the SSX1 mRNA of the peripheral blood was examined in 25 cases of HCC patients and 20 non-HCC patients. The same method was used to detect the expression of SSX-1 mRNA in the tumor tissues, para-tumor tissues, cirrhosis tissues and normal hepatic tissues. A randomized sample was extracted for DNA sequencing from positive electrophoresis expression samples of SSX-1 to examine the reliability of results. ResultsThe expression rates of SSX-1 mRNA were 60%(15/25) and 40%（10/25） respectively in tumor tissues of HCC and the corresponding peripheral blood. SSX-1 mRNA were not expressed in para-tumor tissues,cirrhosis tissues and normal hepatic tissues. The DNA sequence -confirmed that the RTPCR products were true target cDNA. No relationships were found between the expression of SSX-1 gene and clinical characteristics, such as age, sex, tumor size, TNM stage， extent of differentiation and serum AFP level (Pgt;0.05). However, in 33%（3/9） patients with normal serum AFP (lt;20 μg/L), specific expression of SSX1 mRNA was observed. ConclusionHigh specific expression of SSX-1 mRNA is observed in the peripheral blood of patients with HCC, it suggests that applying it as a tumor marker to detect HCC cells in peripheral blood is an adjuvant diagnostic tool. The expression of SSX-1 mRNA in the peripheral blood is observed in the group HCC patients whose serum AFP (lt;20 μg/L) are normal, which suggests that applying both SSX-1 mRNA and AFP as tumor markers together might be useful to improve the diagnostic accuracy for HCC.
ObjectiveTo investigate the clinical significance of CK20 mRNA expression in blood of patients with colorectal cancer. MethodsThe expressions of CK20 mRNA in blood of twenty healthy volunteers, ten patients with colorectal polyp and sixtyone patients with colorectal cancer were detected by RT-PCR. ResultsThe positive rate of CK20 mRNA in peripheral venous blood and portal venous blood of patients with colorectal cancer were 41.0%(25/61) and 45.9%(28/61), which was not significantly different (Pgt;0.05). The expression of CK20 mRNA in patients with colorectal cancer was associated with clinical TNM stage of tumor, local lymph node metastasis, distance metastasis, and the depth of invasion (Plt;0.05). No expression of CK20 mRNA was detected in blood of twenty healthy volunteer’s and ten patients with colorectal polyp. ConclusionCK20 is a specific marker for detecting blood micrometastasis of colorectal cancer. The expression of CK20 mRNA in blood of patients with colorectal cancer is related with TNM stage, invasion, and metastasis of colorectal cancer.
Objective To investigate the expression of ADAM9 in breast cancer and its clinical significance. Methods The expressions of ADAM9 in normal breast tissues and breast cancer tissues were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry, and whose relationship with clinicopathologic features was analyzed. Results The expression of ADAM9 mRNA increased in the breast cancer tissues, but which was not detected in the normal breast tissues. The expression of ADAM9 protein in the breast cancer tissues was significantly higher than that in the normal breast tissues (Plt;0.05), and which in the metastatic lymph nodes was significantly higher than that in the negative lymph nodes or corresponding primary lesions (Plt;0.05). The expression of ADAM9 in the breast cancer tissues was correlated with the lymph node metastasis and histological grade (Plt;0.05). Conclusion ADAM9 is overexpressed in the breast cancer tissues, which might involve in the pathological progression of breast cancer.