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 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.
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 investigate the operative method and to evaluate the cl inical outcome of proximal femoral nail antirotation (PFNA) in treating reverse obl ique fractures of intertrochanteric region of the femur. Methods From January 2007 to February 2008, 30 cases of reverse obl ique fractures of intertrochanteric region of the femur were treated by closed reduction and fixation with PFNA, including 14 males and 16 females and aging 40-88 years old with an average of 68.6years old. All patients had closed fractures. According to AO classification, there were 6 cases of 31-A3.1 type, 7 cases of 31-A3.2 type and 17 cases of 31-A3.3 types. The time from injury to operation was 2-14 days (with an average of 5.3 days). All 31-A3.1 and 31-A3.2 type farctures and 9 cases of AO 31-A3.3 type fractures were fixed with the standard PFNA, and 8 cases of 31-A3.3 type fractures with the PFNA-long. The cl inical and radiological examinations were done at 1, 2, 3, 6, 12, and 18 months after operation. The cl inical outcomes were evaluated according to the Sanders scoring. Results Iatrogenic fracture of femoral shaft occurred in 1 case; no additional procedures were appl ied as fracture kept favorable stabil ity. Superficial infection occurred in 1 case at 5 days after operation, wound healed after dressing change and intravenous antibiotic therapy. Other wounds healed by first intention. All cases were followed up for 12-19 months (mean 14.1 months). All fractures healed uneventfully after 12-30 weeks (mean 16.2 weeks). Five patients complained of hip pain, 2 patients had lateral leg pain, and the pain was el iminated after symptomatic treatment. One case compl icated by ipsilateral fracture of the tibial plateau had functional disturbance of knee, and one case compl icated by ipsilateral fracture of the acetabulum and pelvis had functional disturbance of hip, and the function was improved after functional exercise. No compl ications such as cut-out or breakage of the implants occurred. According to Sanders criteria, the cl inical results were excellent in 22 cases, good in 6 cases, and poor in 2 cases. The excellent and good rate was 93.3%. Conclusion PFNA is an effective treatment method for reverse obl ique fractures of intertrochanteric region of the femur, with a high rate of bone union, minor soft tissue damage, early functional exercise and acceptable compl icationrelated to implant.
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.
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)
【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.