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 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)
OBJECTIVE To study the relationship between the changes of mRNA expression in wound tissues of diabetic ulcers and tissue repair. METHODS The mRNA expression of TGF-beta 1 and IL-6 in eight bioptic samples of diabetic ulcers were detected by RT-PCR and pathologic methods, and the surrounding normal skins from the same patients were measured as control group. RESULTS The mRNA expression levels of TGF-beta 1 were markedly decreased in the diabetic ulcers compared with control group, while the mRNA expression levels of IL-6 were increased at the same reaction conditions. CONCLUSION The different changes of mRNA expression level of TGF-beta 1 and IL-6 in wound tissue result in low production and decreased activity of TGF-beta 1 and IL-6, which lower the reparative ability of wound tissue.
ObjectiveTo investigate the expressions and clinical significance of human telomerase reverse transcriptase (hTERT) mRNA and γglutamyl transpeptidase mRNA-H (GGT mRNA-H) in the peripheral blood of small hepatocellular carcinoma (HCC) patients. MethodsThe expressions of hTERT mRNA and GGT mRNA-H were detected in the peripheral blood of thirty patients with small HCC by RT-PCR, eighteen patients with benign liver diseases, and twelve normal volunteers. ResultsThe positive rate of hTERT mRNA and GGT mRNA-H expression in patients with small HCC were 80.0% (24/30) and 46.7%(14/30), respectively. In patients with hepatitic cirrhosis the positive rate of hTERT mRNA expression was 33.3% (6/18), while the expression of GGT mRNA was not detected. Both the expressions of hTERT mRNA and GGT mRNA-H were negative in all normal volunteers. The combination analysis of hTERT mRNA and GGT mRNA-H expression achieved positive rate of 86.7% in the diagnosis of small HCC, which was significantly higher than the positive rate of AFP (26.7%)， Plt;0.05. ConclusionThe hTERT mRNA and GGT mRNA-H are significantly expressed in small HCC patients, the combination analysis of hTERT mRNA and GGT mRNA-H seems to be useful in the early diagnosis of small HCC.
【摘要】 目的 探讨艾滋病患者合并机会性感染的临床特征并评价其治疗效果。 方法 回顾分析2003年-2009年53例确诊为艾滋病患者的临床资料，对患者合并机会性感染的临床特征，包括发生机会性感染的时间、发生机会性感染时CD4+T细胞计数、起病急缓、严重程度等，进行观察和分析，并给予国家推荐的标准治疗方案进行治疗，通过临床症状、病毒载量、CD4+T细胞计数、影像学监测进行疗效分析，随访时间为初治至出院后6年。 结果 53例艾滋病患者均为重症感染，41例为混合感染，其中2个以上部位感染者为36例（67.9%），2种以上病原体感染者为28例（52.8%），3种以上病原体感染者13例（24.5%）。最常见的机会性感染为结核35.8%（19/53）；其次为卡氏肺孢子菌肺炎30.2%（16/53）；败血症20.8%（11/53），此外，尚有隐球菌感染15.1%（8/53）、弓形虫感染3.8%（2/53）、带状疱疹病毒感染7.5%（4/53）、念珠菌感染17.0%（9/53）、巨细胞病毒感染7.5%（4/53）、合并乙肝11.3%（6/53）、丙肝3.8%（2/53）。机会性感染治疗有效率为77.4%（41/53），病死率为22.6%（12/53），其中隐球菌脑膜炎或混合感染者病死率最高。 结论 艾滋病患者在CD4+T细胞计数lt;350/mm3时各种机会感染明显升高，且随着CD4+T细胞计数的下降呈增高趋势，艾滋病合并机会性感染最常见的是结核，其次卡氏肺孢子菌肺炎；合并结核的治疗效果较好，合并隐球菌脑病的病死率最高。早期启动高效抗逆转录病毒治疗效果好。【Abstract】 Objective To investigate the clinical characteristics of acquired immune deficiency syndrome (AIDS) combined with opportunistic infections and its treatment. Methods We retrospectively analyzed the clinical data of 53 patients diagnosed to have AIDS in our hospital between 2003 and 2009. The clinical features (such as time of the onset of opportunistic infections, CD4+ T cells value at the onset, severity of the infection, etc.) of the opportunistic infections (such as tuberculosis, Pneumocystis carinii pneumonia, toxoplasmosis, etc.) were also observed and analyzed. The patients were treated according to the national standards. The clinical outcome was analyzed based on such indicators as clinical symptoms, viral load, CD4+ T cells value at the onset, and the results of various imaging. Follow-up was done for six years. Results All 53 patients had severe AIDS infection. Forty-one of them had combined infections, among whom 36 (67.9%) had infections in two or more parts of the body, 28 (52.8%) were infected by two or more pathogens, and 13 (24.5%) were infected by three or more pathogens. The most common opportunistic infection was tuberculosis (35.8%,19/53) followed by pneumocystis carinii pneumonia (30.2%,16/53) and septicemia (20.8%,11/53). Other infections included cryptococcal infection (15.1%, 8/53), toxoplasma gondii infection (3.8%, 2/53), herpes zoster virus infection (7.5%, 4/53), candidiasis (17.0%, 9/53), cytomegalovirus infection (7.5%, 4/53), combined hepatitis B (11.3%, 6/53), and hepatitis C (3.8%, 2/53). Effective treatment rate for opportunistic infections was 77.4% (41/53) with a mortality of 22.6% (12/53). The highest fatality rate occurred to those patients with cryptococcal meningitis or mixed infections. Conclusions The occurrence of opportunistic infections rises obviously when CD4+ T cells is lower than 350/mm3, and it increases more significantly as the value of CD4+ T cells goes down. The most common opportunistic infection is tuberculosis followed by pneumocystis carinii pneumonia. The clinical outcome for the combined cases of tuberculosis is good, while combined cryptococcal encephalopathy has the highest mortality rate. High active anti-retroviral therapy should be initiated as early as possible.
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.
OBJECTIVE: To construct a plasmid which has a reporter gene for exploring the role of human telomerase reverse transcriptase(hTRT) in in-vitro cell cultivation. METHODS: hTRT was cut by restricted enzyme from plasmid pGRN145 and inserted to plasmid pEGFP-C1 (enhanced green fluorescent protein). RESULTS: Restricted enzyme analysis and DNA sequencing showed that the sequence of the pEGFP -hTRT transgenic plasmid was correct. CONCLUSION: The recombinant vector pEGFP-hTRT has been successfully constructed, and it can be used as a transgenic plasmid in generating immortalized cell lines.
【摘要】 目的 研究过氧化酶增殖因子活化受体γ（peroxisome proliferator-activated receptor gamma， PPARγ）和血管内皮生长因子C（vascular endothelial growth factor C， VEGF-C）在胃癌组织中表达的相关性，分析其表达与临床病理因素之间的关系。 方法 2005年3月-2009年3月间采用逆转录-聚合酶链反应方法检测36例胃癌手术标本中PPARγ和VEGF-C mRNA的表达，同时选取相同患者的胃正常组织作为对照。 结果 PPARγ mRNA在胃癌中的表达量高于癌旁正常组织，两者的差异有统计学意义（P=0.007）；VEGF-C在胃癌中的表达量高于癌旁正常组织，两者的差异有统计学意义（P=0.004）；PPARγ的表达与VEGF-C表达无关联性（r=0.135，P=0.414）；PPARγ表达与胃癌组织中浸润程度有关（χ2=4.620，P=0.032）、淋巴结转移有关（χ2=15.753，P=0.000）和临床病理分期有关（χ2=4.610，P=0.032）；VEGF-C表达与胃癌组织中淋巴结转移有关（χ2=4.729，P=0.030）、远处转移有关（χ2=4.064，P=0.044）和临床病理分期有关（χ2=6.300，P=0.012）。 结论 PPARγ和VEGF-C可能在胃癌新生淋巴管形成中起重要作用，两者的表达水平与胃癌患者的病情判断及预后评价密切相关。【Abstract】 Objective To investigate the significance of expressions of peroxisome proliferator-activated receptor gamma (PPARγ) and vascular endothelial growth factor C (VEGF-C) in gastric carcinoma as well as their correlation, and to study the relationship between the expressions and clinicopathologic characteristics of gastric carcinoma. Methods Thirty-six pairs of normal mucosa and cancer specimens were obtained from patients who had undergone gastric operation for primary gastric carcinoma and subjected to reverse transcription-ploymerase chain reaction (RT-PCR) for PPARγ and VEGF-C mRNA detection. Results The positive rate and level of PPARγ mRNA expression were higher in gastric cancer tissues than those in normal gastric mucosa (P=0.007). The positive rate and level of VEGF-C mRNA expression were also higher in gastric cancer tissues than those in normal gastric mucosa (P=0.004). Simultaneously, the expression of PPARγ was not correlated with that of VEGF-C (r=0.135, P=0.414), while the highly productions of PPARγ and VEGF-C in gastric carcinomas were both associated with the lymph node metastasis and the clinical stage (Plt;0.05). Conclusion PPARγ and VEGF-C may play an important role in the lymphangio-genesis of gastric cancer, and united detection of PPARγ and VEGF-C expressions may be correlated with making diagnosis, evaluating prognosis in patients with gastric cancer at the same time.
Objective To study the significance of c-met mRNA in axillary drainage after operations for breast cancer. Methods RT-PCR assay was used to examine c-met mRNA in axillary drainage after operations in 52 cases of breast cancer. The relationships between the expression of c-met and the tumor size, metastatic lymph nodes， the expressions of estrogen receptor (ER), progesterone receptor (PR) and c-erbB-2 were analyzed, respectively. In addition, the effect of douching operative field with 5-FU and distilled water on the expression of c-met mRNA was also analyzed. Results ①The proto-oncogene c-met mRNA could be detected in axillary drainage after operations for breast cancer by RT-PCR, and its positive rate was higher than that in routine pathological detection for micrometastasis in the axillary lymph nodes (P＜0.05). ②The expression of c-met mRNA was correlated with both the metastatic lymph nodes and tumor size. ③There was no significant relationship between the expression of c-met mRNA and the expressions of ER, PR and c-erbB-2. ④Dounching operative field with 5-FU and distilled water could decrease the expression of c-met mRNA.Conclusion The proto-oncogene c-met mRNA may be an ideal and specific marker for dectecting micrometastasis of breast cancer. In addition, it also suggests that the examination of c-met mRNA in the axillary drainage by RT-PCR assay could detect the micrometastasis in axillary lymph nodes much easier and more accurately than routine pathological method.