【Abstract】Objective To study the relationship of the expression of CD44v6 mRNA and nm23H1 mRNA with the clinical pathology parameter and prognosis of breast cancer, and to investigate the correlation of the expression of CD44v6 mRNA and nm23H1 mRNA. Methods In situ hybridization and CSA immunohistochemistry method were used to detect the expression of CD44v6 mRNA and nm23H1 mRNA in 94 cases of breast cancer. Results The positive expression of CD44v6 mRNA and the negative expression of nm23H1 mRNA were positively correlated with the grading, clinical stage, lymph node metastasis, and recurrence and prognosis of breast cancer. CD44v6 mRNA expression and nm23H1 mRNA were negatively correlated in breast cancer. Patients who had positive expression of CD44v6 mRNA and negative expression of nm23H1 mRNA had a higher lymph node metastatic rate and a lower survival rate. Conclusion Several genes were involved in the occurrence and development of breast cancer in which the expression of CD44v6 mRNA has synergistic action in negative regulation with that of nm23H1 mRNA. Combined detection of the expression of these two mRNA is helpful to judge the metastasis, recurrence and prognosis of breast cancer.
ObjectiveTo study the expression of HOX A9 mRNA and its clinicopathological significance in the benign and malignant lesions of pancreas. MethodsIn situ hybridization for HOX A9 mRNA was used on routine paraffinembedded sections. ResultsThe positive rate and scoring mean of HOX A9 mRNA expression was significanfly lower in pancreatic carcinoma (49%, 3.3±2.1) than that in chronic pancreatitis (95%, 5.4±0.8) and pericancerous tissues (80%, 4.6±1.2), the negative case of HOX A9 mRNA in chronic pancreatitis and pericancerous tissues showed middle or severelyatypical hyperplasis of the ductal epitheli. The positive rate and scoring mean of HOX A9 mRNA expression was significantly higher in the cases of welldifferentiation (63%, 4.0±2.2) or without metastasis (64%, 4.1±2.2) than that in the ones of poorlydifferentiation (32%, 2.6±2.3) or with metastasis (32%, 2.7±2.2). ConclusionThe expression of HOX A9 mRNA might be related the carcinogenesis, progress, biological behaviors, and prognosis of pancreatic carcinoma. The assay of HOX A9 mRNA expression in the benign lesions of pancreas might have important clinical values in the prevention and earlystage finding of the pancreatic carcinoma.
Objective
To investigate the influnce of L-arginine (L-Arg) and L-nitro-arginine-methyl-ester(L-NAME) to purified retinal ganglion cells(RGCs) apoptosis of rats cultured in different consistencies of L-Arg and L-NAME.
Method
RGCs from Sprague Dawley (SD) neonatal rats(postnatal 1~5 day) were cultured in assimilative culture solution in vitro and RGCs were purified by Thy1.1 with sheep anti rat FITC monoclonal antibody. RGCs were cultured in different consistencies of L-Arg and L-NAME: 1×10-6, 1×10-5,1×10-4, 1×10-3, 1×10-2 and 1×10-1 mol/L for 24 hours and 48 hours, respectively. The changes of bcl-2, bax and p53 mRNA in RGCs in different consistencies of L-Arg and L-NAME
were demonstrated qualitatively and quantitatively by in situ hybridization, and their apoptosis were detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) method, respectively.
Results
After 24 hours in vitro, the purification rate of RGCs in the experiment arrived at 97 %. After 48 hours, there were a few apoptotic cells expression in the control group. Apoptotic cells expression in L-Arg≥1×10-3 mol/L and L-NAME≥1×10-1 mol/L groups increased that had a significant difference with the control group (Plt;0.05). In the group of L-Arg≥1×10-3 mol/L and L-NAME≥1×10-1 mol/L, the expression of bcl-2 mRNA in RGCs became weaker and weaker as the consistencies were increased, but the expression of bax and p53 mRNA in RGCs became higher and higher and had a significant difference with control group (Plt;0.05).
Conclusion
Lower concentration of L-Arg can promote the growth of purified RGCs in vitro and higher concentration of L-Arg can promote the apoptosis of RGCs.
(Chin J Ocul Fundus Dis, 2002, 18: 137-139)
Objective
To observe the expression of proinflammatory factors messenger RNA(mRNA) in periretinal membrane of proliferative vitreoretinopathy.
Methods
Fourteen specimens of periretinal membrane were collected during vitrectomy, and they were made into paraffin sections.The presence of mRNA coding for IL-1,IL-6,IL-8 and TNF alpha was observed by in situ hybridization(ISH) with biotin labeled oligonuclotide probes respectively.The eyeball after corneal grafting was made as normal control.
Results
No expression of proinflammatory factors mRNA was found in normal human retina.Positive staining was present in 5 specimens.In these specimens, IL-1βmRNA was found in 3 specimens and TNFαmRNA was found in 3 specimens,there is 1 specimen expressed IL-8 mRNA and 3 specimens expressed IL-6 mRNA.In these positive specimens, one contained cells expressing mRNA for IL-1βbeta and IL-6, and one exhibited cells expressing mRNA for IL-1β、IL-8 and TNFα,two membranes possessed positive cells for IL-6 and TNFαmRNA, one membrane contained IL-1βmRNA positive cells only.
Conclusion
These findings suggest that these cytokines may be locally produced by cells infiltrating epiretinal membranes. The expression of IL-1β, IL-6, IL-8 and TNFαmRNA within retinal membranes provides further evidence of a pathogenic role of these cytokines in proliferative vitreoretinopathy.
(Chin J Ocul Fundus Dis, 2002, 18: 286-288)
Objective To investigate the association of the expression of CD15 mRNA with the invasion and prognosis of hepatocellular carcinoma (HCC) and the expression of nm23H1 mRNA. Methods In situ hybridization and immunohistochemistry methods were used to detect the expression of CD15 mRNA and protein nm23H1 mRNA in HCC.Results In 99 cases of HCC, the positive rate of CD15 mRNA,its protein and nm23H1 mRNA were 38.4%, 36.4% and 76.8%, respectively. The expression of CD15 mRNA was consistent with its protein and negatively correlated with the expression of nm23H1 mRNA. The expression of CD15 mRNA and its protein, nm23H1 mRNA were associated with the invasiveness and metastasis of HCC and the prognosis of HCC patients. Conclusion The detection of CD15 expression could be a new pathological biology index to judge the metastasis and prognosis of HCC.
Objective Using molecular biology method to detect and genotype human papilloma virus (HPV) in women taking physical examination in West China Hospital, Sichuan University, to explore the infection status and genotype distribution of HPV in normal women in Chengdu area, and to provide basis for early effective prevention and control of cervical cancer and domestic research and development of HPV vaccine. Methods Flow fluorescent hybridization technique was used to detect and genotype HPV-DNA in 25 148 healthy women taking physical examination in West China Hospital, Sichuan University between May 1st, 2018 and May 31st, 2019. The overall positive HPV infection rate, HPV genotype distribution, and characteristics of HPV infections were analyzed and calculated, and the HPV infection rates of different age groups were calculated and compared by chi-square test using SPSS 17.0 software. Results The overall positive rate of HPV infection was 12.19% (3 066/25 148). The high-risk HPV genotypes infection rate was 8.69% (2 186/25 148), and the top five subtypes with the highest infection rates were HPV52, HPV53, HPV58, HPV16, and HPV39. The low-risk HPV genotypes infection rate was 4.66% (1 171/25 148), and the top five subtypes with the highest infection rates were HPV61, HPV81, HPV43, HPV44, and HPV6. Single subtype infections were the main infections with a proportion of 81.74% (2 506/3 066), and the most common multiple infections were double infections which accounted for 13.96% (428/3 066). In different age groups, the HPV infection rate of group 60-69 was the highest (12.87%), while that of group 70-89 was the lowest (10.88%), but the difference among different age groups was not statistically significant (χ2=4.035, P=0.544). Conclusion According to the results of this study in women taking physical examination in West China Hospital, Sichuan University, we suggest adding HPV52, HPV53, and HPV58 which have the highest infection rate in high-risk HPV subtypes to the evaluation of domestic HPV vaccine screening and the cervical cancer prevention and control system.
ObjectiveTo investigate the expression and relation of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in rats with diabetic retinopathy.MethodFifty-five Wistar rats were randomly divided into the control group (10 rats), and 1, 3, and 5-month-diabetes group (15 rats in each diabetes group), and the diabetic models were set up. The expressions of VEGF and bFGF were detected by situ hybridation and immunohistochemistry on retinal paraffin sections.ResultsThe results of situ hybridation showed that expression of bFGF was found in 3-month-deatbtes group with the percentage of 77.8%, and 88.9% in 5-month-deatbtes group; the positive expression of VEGF was not found in 3-month-deatbtes group but in 5-month-deatbtes group with the percentage of 66.7%. Immunohistochemistry indicated that the positive expression of bFGF started in 3-month-deatbtes group with the percentage of 55.6%, and 88.9% in 5-month-deatbtes group; the percentage of the expression of VEGF was 33.3% in 3-month-deatbtes group and 88.9% in 5-month-deatbtes group.ConclusionThe expression of VEGF occurs after the expression of bFGF in rats with DR.(Chin J Ocul Fundus Dis, 2005,21:37-40)
Objective To identify and isolate the variant gene associated with gastric adenocarcinoma and clone the fragment of variant gene.Methods By arbitrarily primer polymerase chain reaction (AP-PCR), DNA samples from 5 matched gastric adenocarcinoma and non-tumor gastric tissues were analysed. Results The produced AP-PCR profiles were different in each matched gastric adenocarcinoma and non-tumor gastric tissue. One differentiated amplified DNA fragments PW2.2 from a matched gastric adenocarcinoma were cloned. The result of Southern blot hybridization with PW2.2 as a probe showing that this fragment was also found in some other gastric adenocarcinoma samples. Conclusion AP-PCR fingerprinting assay can be used to identify and clone the variant genes associated with gastric adenocarcinoma.
Objective To identify the expression of pleiotrophin （PTN） mRNA and protein in the colorectal cancer tissues， and to explore the clinical value of it. Methods The expressions of PTN mRNA and protein in colorectal cancer tissues （colorectal cancer group） as well as normal colorectal tissues （normal control group） were tested by using in-situ hybridization and immunohistochemistry. Results The positive rates of PTN mRNA 〔63.9% （53/83） vs. 40.7%（22/54）〕 and protein〔60.2%（50/83） vs. 33.3%（18/54）〕 in colorectal cancer group were all significantly higher than those of normal control group （P=0.008， P=0.002）. There were no significant relationship between expressions of PTN mRNA and protein with gender， age， and type of tumor （P＞0.05）， but in tissues of Ⅲ-Ⅳ stage and poor differentiation，the positive rates of PTN mRNA and protein were all higher （P＜0.05）. Conclusions The over expressions of PTN mRNA and protein in colorectal cancer tissues may directly related to the invasion and metastasis. Meanwhile， it can be used as an index to predict metastasis and prognosis of colorectal cancer.
ObjectiveTo observe the expression of connective tissue growth factor (CTGF) in injured model of retinal pigment epithelial (RPE) cells and the promoting effect of CTGF on migration of RPE cells.MethodsCultured monolayer-confluent human RPE cells were scraped with a trephine and a cotton stick, and set up the injured model of RPE cells with round scraped area. Immunohistochemistry and in situ hybridization(ISH) were used to detect the expression of CTGF protein and mRNA in injured RPE cells at distinct time points after injury. The number of RPE cells migrated to injured area was measured and the effect of CTGF on migration of RPE cells and the effect of dexamethasone (DEX) on the promoting process of CTGF were observed.ResultsThe results of immunohstochemistry and ISH indicated the weak positive expression of CTGF in RPE cells at the edge of scrape 6 hours after injury, and the positive expression increased gradually as time goes by after the injury. Strong positive expression of CTGF in RPE cells at the edge of scrape was found 24 and 48 hours after injury. Rebuilt human CTGF stimulated migration of RPE cells in a dose-depended manner, and DEX significantly inhabited the migration.ConclusionCTGF involves in the procedure of repair of injury of RPE cells, which may play an important role in the pathogenesis of intraocular proliferative diseases such as proliferative vitreoretinaopathy.(Chin J Ocul Fundus Dis, 2005,21:306-309)
