ObjectiveTo evaluate the possible role of the expression of insulin-like growth factor-1 receptor (IGF-1R) in determining rectal cancer radiosensitivity. MethodsThe paired preradiation biopsy specimens and postoperative specimens were obtained from 87 patients with rectal cancer in the department of digestive tumor surgery, Jiangsu Province Hospital of Traditional Chinese Medicine, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine from January 2009 to December 2010. The IGF-1R expression was examined by immunohistochemistry (IHC) and reverse transcription-polymerase chain reaction (RT-PCR). The tumor radiosensitivity was defined according to Rectal Cancer Regression Grade, then the relation between the IGF-1R expression and tumor radiosensitivity was evaluated. ResultsCompared with the preradiation biopsy specimens, IGF-1R expression significantly increased in the paired postoperative specimens of the residual cancer cells (Plt;0.001). The IHC result demonstrated IGF-1R overexpression was significantly associated with a poor response to radiotherapy (rs=0.401, Plt;0.001); RT-PCR detection of IGF-1R expression on preradiation biopsy specimens also showed that IGF-1R mRNA negative patients had a higher radiation sensitivity (rs=0.497, Plt;0.001). ConclusionDetection of IGF-1R expression may predict radiosensitivity of preoperative irradiation for rectal cancer.
Objective To investigate the effect of hypoxia on expressions of erythropoietin（EPO）mRNA and protein in retinal Muuml;ller cells cultured in vitro. Methods Retina tissues from the new-born Wistar rats were dissected into cell suspension after digested by pancreatin.Muuml;ller cells were separated and purified by mechanical concussion and blowing and striking method.The expression of EPO mRNA and protein under the condition of hypoxia was detected by semi-quantitative reverse transcriptase（RT）-polymerase chain reaction（PCR）and immunocytochemical method. Results Retinal Muuml;ller cells were cultured successfully，95% of which were positively stained by glial fibrillary acidic protein（GFAP）.Positively stained EPO protein was located in the cytoplasm and protuberance.The expression of EPO mRNA and protein was faint in the normal retinal Muuml;ller cells，but increased obviously and time-dependently after hypoxia. Conclusion Expression of EPO mRNA and protein increases in Muuml;ller cells after hypoxia，which may be one of the protective factors for the nerves in anoxic retinopathy. (Chin J Ocul Fundus Dis, 2006, 22: 196-199)
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.
Objective：To observe the expression of gene and protein l evel of unfolded protein, glucoseregulated protein 78 (GRP78), after retinal d etachment (RD); to find out the relationship between UPR and the cell damage after RD. Methods：Eightyeight Wistar rats were divided into 2 groups: con trol group (11 rats) and RD group (77 rats). In RD group, subretinal injection with 10 mg/ml hyaluronic acid sodium was performed on the left eyes of the rats t o set up RD model, and the left eyes and retinal tissue were collected 1/2 day, 1 day, 2, 4, 8, 1 6 and 32 days after RD; there were 11 rats in each subgroup. The expression of G RP78 mRNA in retina tissue was detected by semiquantitative reverse transcript i on polymerase chain reaction (RT－PCR), the expression of GRP78 protein level wa s detected by Western blotting, and the distribution of GRP78 in each retinal lay er was observed by immunofluorescence labeling method and confocal microscopy. Results：The expression of retinal GRP78 mRNA significantly in creased in 1/2 day , 1 day, 2, and 4 days subgroups after RD (Plt;0.05). The expression of GRP7 8 protein significantly increased in each subgroup after RD compared with which in the control group, and reached the peak in 8, 16, and 32 days subgroups. The expres sion of GRP78 protein was detected in all of the retinal layers after RD. Conclusion：The protection mechanism of UPR starts up after RD, and l eads the correc t pucker of the protein and reduces cellular injury by upregulating the expres s ion of GRP78, which provide the theoretic basis for reducing the cellular injury and improving the visual function in patients with RD.
Objective To discuss the changes of c-kit/scf mRNA and protein in guinea pig gallbladder fed on high cholesterol diet. Methods Twenty guinea pigs were divided into two equal groups of 10 each：the control group and lithogenic group. Normal diet and high cholesterol diet was given to each group respectively. The period of stone permeation was six weeks. RT-PCR and Western blot were used to determin the expressions of c-kit and scf mRNA and protein. Results RT-PCR results showed that the expressions of c-kit mRNA（t=6.985，P＜0.01） and scf mRNA （t=6.028， P＜0.01）decreased significantly in lithogenic group compared with the control group. Western blot results showed that the expressions of c-kit protein （t=10.256， P＜0.01） and scf protein （t=9.586， P＜0.01）decreased significantly in lithogenic group compared with the control group. Conclusions The expressions of c-kit/scf mRNA and protein decrease during the formation of cholesterol gallstones in guinea pigs fed on high cholesterol diet. Inhibition of c-kit/scf pathway may play a role in the formation of cholesterol gallstones.
Objective To detect the expression of forkhead box P3 （FOXP3 ）gene in esophageal squamous cell carcinoma(ESCC) and provide a new basis for immunotherapy of esophageal cancer. Methods Based on fluorescent TaqMan methodology, a realtime quantitative reverse transcription polymerase chain reaction (RT-PCR) for detecting the expression of FOXP3 was set up. In this method, a cloning vector pMD 18-T-FOXP3 was constructed as a standard plasmid. The specific expression of FOXP3 in 42 patients with ESCC and 30 healthy controls were measured by using GeneAmp 7500 Sequence Detection Systems. Results FOXP3 mRNA copy number in ESCC was significantly higher than that in healthy control tissue ［(72.20±23.10)×104copy/μg RNA vs.(0.68±0.34)×104 copy/μg RNA；Plt;0.05］. Conclusion A realtime quantitative RT-PCR method for detecting the expression of FOXP3 gene in ESCC has been successfully established. The expression level of FOXP3 is increased in ESCC compare with healthy controls.
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.