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The cultivation of medical students' scientific research and innovation ability is the need of our country to accelerate the construction of an innovative country. This paper starts from the significance of strengthening the cultivation of medical students' scientific research and innovation ability, emphasizes participation in teacher's scientific research in colleges and universities to strengthen the feasibility of the medical students' scientific research and innovation ability cultivation, and summarizes the practical experience of cultivating medical students' scientific research ability based on scientific research projects of teachers in colleges and universities, which can provide reference for peers.
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Objective :To analyze therapeutic effect of early standardized swallowing rehabilitation training combined acupuncture on patients with dysphagia after stroke .Methods :A total of 98 patients with dysphagia after stroke were selected ,randomly and equally divided into swallowing training group (received early standardized swallowing rehabilitation training) and combined treatment group (received acupuncture based on swallowing training group ). All patients were assessed by Scores of Kubota＇s water drinking test and Ichiro Fujishima dysphagia score and its re‐ sults were compared between two groups before and three months after treatment .Results :Compared with before treatment ,there was significant reduction in score of Kubota＇s water drinking test ,and significant rise in Ichiro Fu‐jishima dysphagia score after three‐month treatment in two groups , P＝0.001 all .Compared with swallowing train‐ing group after three‐month treatment , there was significant reduction in score of Kubota＇s water drinking test [ (1.83 ± 0.74) scores vs.(1. 41 ± 0. 63) scores] ,and significant rise in Ichiro Fujishima dysphagia score [ (8. 96 ± 1.23) scores vs.(9. 86 ± 1. 04) scores] in combined treatment group , P＜0.01 both .Dysphagia recover‐to‐normal rate of combined treatment group was significantly higher than that of swallowing training group (69.39% vs. 42. 6%, P＝ 0. 008 ).Conclusion : Early standardized swallowing rehabilitation training combined acupuncture can help to improve Clin symptoms and enhance therapeutic effect in patients with dysphagia after stroke .
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Objective To preliminarily test the value of gastroesophageal reflux disease questionnaire (GERD-Q) in auxiliary diagnosis of gastroesophageal reflux disease(GERD) in infants and young children.Methods From January 2011 to February 2014,148 infants and 87 young children with suspected GERD symptoms were enrolled in the outpatient and inpatient departments of Wuhan Women and Children's Medical Care Center.The diagnosis of GERD were made through 24 h esophageal pH monitoring and upper gastrointestinal hysterosalpingography.Those with any examinations being positive were brought into GERD group,and the others into the control group.All cases had been surveyed with GERD-Q.Using SPSS17.0 statistical software,the ratio was compared with chi-square test,and the difference of the scores between groups with t test.Results In infants (1 to 11 months),the individual symptom scores (ISS) of the 6 symptoms in the GERD group (n =111) were all higher than those in the control group (37 cases) (P ＜ 0.05),and the composite and symptom scores (CSS) [(131.43 ± 66.56) scores] in the GERD group was higher than that [(33.70 ± 40.03) scores] in the control group (t =8.42,P ＜ 0.05).In young children (1 to 3 years old),the ISS in the GERD group(55 cases) were all higher than those in the control group(32 cases),and the CSS[(98.58 ± 66.75) scores] in the GERD group was also higher than that [(28.50 ± 35.95) scores] in the control group(t =5.49,P ＜ 0.05).Conclusions There is a good correlation for age-specific GERD-Q to distinguish GERD symptoms in infants and young children,which is suitable for the assessment of GERD.
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The present study aimed to compare the clinical efficacy of a 0.4% carbomer gel and 1% carboxymethyl cellulose (CMC) containing artificial tears in treatment of dry eye patients. Sixty subjects with mean age of 45.89 years who had symptoms and signs of dry eye were enrolled in this prospective, investigator-masked and stratified random sampling study. The subjects were divided into two parallel groups with 30 subjects (60 eyes) in each group. One group received carbomer gel, and the other group received 1% CMC containing artificial tears. Subjects received the drops 3 to 4 times or more per day for 3 months. At the first visit time, the precorneal residence time of these two drops was measured. The efficacy was assessed by comparing the subjective symptoms (ocular dryness, foreign body sensation, burning sensation and pain), and the objective test results of tears breakup time, Schirmer's test and corneal fluorescein staining prior to the study and after the treatment. As a result, the ocular residence time of carbomer gel was significantly longer than that of 1% CMC (P<0.001). Most of the primary subjective symptoms and objective test results were improved after treatment in both carbomer gel group and 1% CMC group. As to the improvement of each symptom and objective test result, carbomer gel was more effective than 1% CMC group (P<0.01). In conclusion, carbomer gel had longer precorneal residence time and was more effective than 1% CMC in the treatment of patients with dry eyes.
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Resinas Acrílicas/administração & dosagem , Carboximetilcelulose Sódica/administração & dosagem , Síndromes do Olho Seco/tratamento farmacológico , Géis/administração & dosagem , Soluções Oftálmicas/administração & dosagem , Estudos ProspectivosRESUMO
In order to construct a recombinant plasmid containing short hairpin RNA (shRNA) targeting survivin and to investigate its effect on survivin expression and cell apoptosis of human retinoblastoma cell line Hxo-rb44 in vitro, RNA interference plasmid pSIRENS that can express shRNA of survivin was designed, constructed, and transfected into human retinoblastoma cell line Hxo-rb44.Survivin and c-Myc expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Apoptosis of Hxo-rb44 cells was assayed by Honchest33258 staining and cell growth curve was drawn. The results showed that the oligonucleotide targeting survivin was identified in pSIRENS plasmid. After pSIRENS plasmid transfected, survivin and c-Myc expression in Hxo-rb44 cells was decreased significantly. Apoptotic rate of cells was up-regulated from (3.5±1.29) % to (36.1±19.66) %. The proliferation ability of Hxo-rb44 cells was inhibited. No significant effects on survivin expression and apoptosis of the cells were found when negative control plasmid was transfected. In conclusion, the plasmid containing shRNA targeting survivin was constructed successfully. It could inhibit efficiently the expression of survivin and c-Myc in human retinoblastoma cell Hxo-rb44 in vitro. The inhibition of the expression of c-Myc might be involved in the apoptosis of Hxo-rb44 cells.
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To evaluate the efficacy and safety of resection and cryotherapy combined with amniotic membrane transplantation (AMT) for the treatment of vernal keratoconjunctivitis (VKC) with giant papillae (GP). Eight patients (16 eyes involved) with VKC, characterized by GP on the upper tarsal conjunctiva, underwent resection and cryotherapy in combination with AMT. The follow-up lasted for 3-22 months. The results showed that corneal shield ulcers and superficial punctuate keratitis healed during the first week after surgery and did not recur. Fourteen eyes (87.5 %) were symptom-free 1 month after surgery, and no GP, ectropion, trichiasis and other complications were noted, but the blood vessels of upper tarsal conjunctiva could not be clearly seen and a little conjunctival scar was observed. Recurrence of GP was observed in 2 eyes (12.5 %), with the area being less and irritation milder as compared with those before the operation. Among the two eyes, one eye was treated by cyclosporine eyedrops with improvement, but the other eye showed no improvement after the treatment, and underwent a second surgery with a cotton patch soaked in fluorouracil applied onto the supratarsal area after resection and cryotherapy. Four months after the treatment the patient presented no symptoms and GP did not recur. It is concluded that the resection and cryotherapy combined with AMT is an effective and safe treatment for VKC with GP.
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The central corneal thickness (CCT) in age 48 years or less of Chinese was characterized and its relationship with gender, age, refraction and intraocular pressure (IOP) was investigated.Right eyes of 1669 participants were included in this study (880 men, 52.7 % and 789 women,47.3 %). Mean age of the samples was 23.8±5.9 years. After the examination of corneal topography and refraction, Goldman applanation tonometry was carried out by one physician. Tonometric values were the mean of three consecutive readings. Subsequently, another physician carried out ultrasonic pachymetry with the DGH 2000 AP ultrasonic pachymeter. Six measuremen ts were made at the center of the cornea of each eye. The mean value was used for analysis. The results showed that mean CCT of male participants was 551.33± 34. 62 μm, 5.79 μm more than that of female participants. Linear regression analyses revealed that CCT was negatively related with age only in female and no association was found between refractive status and CCT. IOP was positively related to CCT, and there was a difference in IOP of 1.5 mmHg (1 mmHg=0. 133 kPa) per 100 μm difference in CCT. Ocular hypertension group was prone to have thicker cornea than average. The results indicated that in adult Chinese CCT tended to decrease with aging in female only. IOP measured by Goldmann tonometry was positively related with CCT so that CCT should be measured along with IOP.
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To study corneal lymphangiogenesis after corneal transplantation, corneal allogenic transplantation models were established in rats. 8 female Wister rats were used as donors, and 16 Sprague Dawley (SD) rats were used as recipients and 2 SD served as controls. Corneal lymphangiogenesis and hemangiogenesis was examined by electron microscopy 1 and 2 weeks after corneal penetrating transplantation, and the expression of lymphatic vessel endothelial receptor (LYVE-1) was examined 1, 3, 7, 14 days after the transplantation respectively. In addition, 19 allograft failed human corneas were examined by 5'-nase-alkaline phosphatase (5'-NA-ALP) doubleenzyme-histochemistry staining to detect corneal lymphangiogenesis and hemangiogenesis. By immunohistochemistry for LYVE-1, it was found that blown lymphatics were localized in the stroma 3days after the corneal transplantation. With electron microscopy, new lymphatic vessels and blood vessels were found 1 and 2 weeks after the corneal transplantation. By 5'-NA-ALP enzyme-histochemistry, corneal hemangiogenesis was found in all allograft failed huma n corneas and 5 of 19(26.3%) cases had developed corneal lymphangiogenesis. It is concluded that corneal lymphangiogenesis is present after corneal transplantation, which may play an important role in allograft rejection.
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In order to investigate the effects of mouse CTLA4Ig gene-modified dendritic cells (DCs) on the survival of the corneal allografts in rats, the plasmid PG\CTLA4Ig was transfected into DCs of F344 rats mediated by LipofectamineTM 2000. The expression of CTLA4Ig was detected by immunofluorescent microscopy. The effects of donor DCs on the proliferation of T cells in Lewis rats (recipients) were tested by by CCK8. Corneal transplantation was performed from F344 rats to Lewis rats. The DCs modified with CTLA4Ig gene were injected into the Lewis rats on the day 0 and 3 after transplantation. The movement of the DCs after modification in vivo was observed by immunofluorescent microscopy, and the survival of corneal allografts was evaluated by Holland criterion. The results showed that the CTLA4Ig-modified DCs could restrain the proliferation of allogenetic T cells. The CTLA4Ig-modified DCs prolonged survival of corneal allografts. (P＜0.01). It was suggested that the injection of CTLA4Ig gene-modified DCs could obviously inhabit the allograft rejection and prolong the survival of corneal allografts.
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To study corneal lymphangiogenesis after corneal transplantation, corneal allogenic transplantation models were established in rats. 8 female Wister rats were used as donors, and 16 Sprague Dawley (SD) rats were used as recipients and 2 SD served as controls. Corneal lymphangiogenesis and hemangiogenesis was examined by electron microscopy 1 and 2 weeks after corneal penetrating transplantation, and the expression of lymphatic vessel endothelial receptor (LYVE-1) was examined 1, 3, 7, 14 days after the transplantation respectively. In addition, 19 allograft failed human corneas were examined by 5'-nase-alkaline phosphatase (5'-NA-ALP) double-enzyme-histochemistry staining to detect corneal lymphangiogenesis and hemangiogenesis. By immunohistochemistry for LYVE-1, it was found that blown lymphatics were localized in the stroma 3 days after the corneal transplantation. With electron microscopy, new lymphatic vessels and blood vessels were found 1 and 2 weeks after the corneal transplantation. By 5'-NA-ALP enzyme-histochemistry, corneal hemangiogenesis was found in all allograft failed human corneas and 5 of 19 (26.3 %) cases had developed corneal lymphangiogenesis. It is concluded that corneal lymphangiogenesis is present after corneal transplantation, which may play an important role in allograft rejection.
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Córnea/irrigação sanguínea , Córnea/química , Córnea/ultraestrutura , Neovascularização da Córnea/etiologia , Neovascularização da Córnea/metabolismo , Transplante de Córnea/efeitos adversos , Transplante de Córnea/métodos , Imuno-Histoquímica , Linfangiogênese , Microscopia Eletrônica , Ratos Sprague-Dawley , Ratos Wistar , Proteínas de Transporte Vesicular/biossínteseRESUMO
To evaluate the efficacy and safety of resection and cryotherapy combined with amniotic membrane transplantation (AMT) for the treatment of vernal keratoconjunctivitis (VKC) with giant papillae (GP). Eight patients (16 eyes involved) with VKC, characterized by GP on the upper tarsal conjunctiva, underwent resection and cryotherapy in combination with AMT. The follow-up lasted for 3-22 months. The results showed that corneal shield ulcers and superficial punctuate keratitis healed during the first week after surgery and did not recur. Fourteen eyes (87.5 %) were symptom-free 1 month after surgery, and no GP, ectropion, trichiasis and other complications were noted, but the blood vessels of upper tarsal conjunctiva could not be clearly seen and a little conjunctival scar was observed. Recurrence of GP was observed in 2 eyes (12.5 %), with the area being less and irritation milder as compared with those before the operation. Among the two eyes, one eye was treated by cyclosporine eyedrops with improvement, but the other eye showed no improvement after the treatment, and underwent a second surgery with a cotton patch soaked in fluorouracil applied onto the supratarsal area after resection and cryotherapy. Four months after the treatment the patient presented no symptoms and GP did not recur. It is concluded that the resection and cryotherapy combined with AMT is an effective and safe treatment for VKC with GP.
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In order to construct a recombinant plasmid containing short hairpin RNA (shRNA) targeting survivin and to investigate its effect on survivin expression and cell apoptosis of human retinoblastoma cell line Hxo-rb44 in vitro, RNA interference plasmid pSIRENS that can express shRNA of survivin was designed, constructed, and transfected into human retinoblastoma cell line Hxo-rb44. Survivin and c-Myc expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Apoptosis of Hxo-rb44 cells was assayed by Honchest33258 staining and cell growth curve was drawn. The results showed that the oligonucleotide targeting survivin was identified in pSIRENS plasmid. After pSIRENS plasmid transfected, survivin and c-Myc expression in Hxo-rb44 cells was decreased significantly. Apoptotic rate of cells was up-regulated from (3.5+/-1.29) % to (36.1+/-19.66) %. The proliferation ability of Hxo-rb44 cells was inhibited. No significant effects on survivin expression and apoptosis of the cells were found when negative control plasmid was transfected. In conclusion, the plasmid containing shRNA targeting survivin was constructed successfully. It could inhibit efficiently the expression of survivin and c-Myc in human retinoblastoma cell Hxo-rb44 in vitro. The inhibition of the expression of c-Myc might be involved in the apoptosis of Hxo-rb44 cells.
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In order to investigate the effects of mouse CTLA4Ig gene-modified dendritic cells (DCs) on the survival of the corneal allografts in rats, the plasmid PG\CTLA4Ig was transfected into DCs of F344 rats mediated by Lipofectamine 2000. The expression of CTLA4Ig was detected by immunofluorescent microscopy. The effects of donor DCs on the proliferation of T cells in Lewis rats (recipients) were tested by by CCK8. Corneal transplantation was performed from F344 rats to Lewis rats. The DCs modified with CTLA4Ig gene were injected into the Lewis rats on the day 0 and 3 after transplantation. The movement of the DCs after modification in vivo was observed by immunofluorescent microscopy, and the survival of corneal allografts was evaluated by Holland criterion. The results showed that the CTLA4Ig-modified DCs could restrain the proliferation of allogenetic T cells. The CTLA4Ig-modified DCs prolonged survival of corneal allografts. (P < 0.01). It was suggested that the injection of CTLA4Ig gene-modified DCs could obviously inhabit the allograft rejection and prolong the survival of corneal allografts.
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The central corneal thickness (CCT) in age 48 years or less of Chinese was characterized and its relationship with gender, age, refraction and intraocular pressure (IOP) was investigated. Right eyes of 1669 participants were included in this study (880 men, 52.7% and 789 women, 47.3%). Mean age of the samples was 23.8 +/- 5.9 years. After the examination of corneal topography and refraction, Goldman applanation tonometry was carried out by one physician. Tonometric values were the mean of three consecutive readings. Subsequently, another physician carried out ultrasonic pachymetry with the DGH 2000 AP ultrasonic pachymeter. Six measurements were made at the center of the cornea of each eye. The mean value was used for analysis. The results showed that mean CCT of male participants was 551.33 +/- 34.62 microm, 5.79 microm more than that of female participants. Linear regression analyses revealed that CCT was negatively related with age only in female and no association was found between refractive status and CCT. IOP was positively related to CCT, and there was a difference in IOP of 1.5 mmHg (1 mmHg = 0.133 kPa) per 100 microm difference in CCT. Ocular hypertension group was prone to have thicker cornea than average. The results indicated that in adult Chinese CCT tended to decrease with aging in female only. IOP measured by Goldmann tonometry was positively related with CCT so that CCT should be measured along with IOP.
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Fatores Etários , China , Córnea/anatomia & histologia , Topografia da Córnea , Pressão Intraocular , Valores de Referência , Fatores Sexuais , Tonometria OcularRESUMO
In order to investigate the ipsilateral lymphadenectomy for inhibiting rejection in rat corneal transplantation, corneal allogenic transplantation models were established in rats. Eighteen female Wister rats were used as donors, and 36 Sprague Dawley rats as recipients. After penetrating corneal transplantation, recipients were randomly divided into 3 groups: group A (control group); group B, the ipsilateral lymphadenectomy group; group C, the bilateral lymphadenectomy group. Among 12 rats in each group, the corneas of 2 rats in each group were used for pathological study at day 14 after the transplantation, and the remaining 10 rats were used for studying corneal rejection by a slit lamp. The time points when allograft rejection occurred were recorded and mean survival time (MST) was compared. The results showed that MST in groups B and C was 46.30 +/- 9. 464 days and 44.43 +/- 7. 604 days, respectively, which was significantly prolonged as compared with that in group A (10.71 +/- 1.567 days, P 0.05). It was concluded that both bilateral and ipsilateral lymphadenectomy therapies could effectively inhibit the corneal allograft rejection. Ipsilateral lymphadenectomy is a less complex surgical procedure and is just as effective in preventing rejection.
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Transplante de Córnea , Rejeição de Enxerto/prevenção & controle , Sobrevivência de Enxerto , Excisão de Linfonodo/métodos , Distribuição Aleatória , Ratos Sprague-Dawley , Ratos WistarRESUMO
BACKGROUND: The expression of main histocompatibility antigen in cornea of rats is similar to that of human. It is verified in early experiments that penetrating corneal transplantation in rats is more repetition and reliability, and the operation difficulty is lower than that in mice.OBJECTIVE: To establish rat model of penetrating corneal transplantation and analyze the reasons of complications during model preparation and the corresponding managements and probe into the method for improving the success rate of rat model of corneal transplantation.DESIGN: Randomized controlled experiment was designed.SETTING: Department of Ophthalmology , Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Department of Ophthalmology in Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from March to May 2004. Seventysix outbred female Wistar rats of two months old were employed and randomized into experimental group and control group with 38 rats in each,without eye disease, of clean grade, body mass varied from 180 to 200 g and their right eyes were taken as acceptors. Thirty-eight SD female rats were employed and their both eyes were taken as donors. All of the animals were provided from Experimental Animal Center of Tongji Medical College of Huazhong University of Science and Technology.METHODS: Central penetrating corneal transplantation in situ was adopted and the model was established on the right eye of Wistar rat. Ten minutes before operation, tropicamide eye drops was administrated twice to amplify adequately the pupil of the right eye up to 4.0 mm. According to body mass, general anesthesia was done with abdominal injection of 3.0 mL/kg chloral hydrateand dicaine eye drops was applied twice 5 minutes before the operation with 0.1 volume fraction. Routine sterilization was done on the head of rat, the eyes kept in horizontal level and respiration kept smooth. The clip in dental-ophthalmology department was used to fix the posterior of the operated eye and make it to be semiluxated. The ring driller 3.5 mm in diameter was used to collect corneas of both eyes in SD rat as the implant tissue and place in culture dish with endothelial part upward, covered with methyl cellulose of 20 g/L mass weight concentration. The ring driller, 3.0 mm in diameter was used to collect the cornea of the right eye in Wistar rat to be the graft foreman, and the implant tissue was sutured into the graft foreman on the right eye of Wistar rat with 10-0nylon thread intermittently. Eight stitches were done in corneal transplantation in the experimental group and 6 stitches in the control group. After operation, ofloxacin eye drops was administrated and the operated eye was covered with plaster. Corneal rejection was observed and the evaluation was done in 3 items, named, turbidity, edema and neogenetic vessel. The total score of 3 items was taken as the rejection index on the day. Rejection was determined if the rejection index on the day ≥5 or corneal turbidity reached the 3nd grade. x2 and Fisher definite probability methods were used for statistical examination.plant tissue and corneal rejection.RESULTS: Of 76 Wistar rat acceptors, 5 rats were dropped out, among which, 1 rat was died accidentally after operation in the experimental group, 2 rats were died accidentally after operation in the control and 2 rats were injured cornea on the operated eyes due to fighting. But by supcomplications after corneal transplantation: In the experimental group, the number with anterior adhesion of sclera, non-round pupil; limited wound healing and non-formation of anterior chamber was lower than that in the implant tissue and corneal rejection: the survival time of corneal implant tissue was similar basically in both experimental group and the control [(8.9±2.3), (8.6±2.3) days, P ＞ 0.05]. Rejection presented in both groups in 16 days after corneal transplantation and the incidence of rejection was 100%.CONCLUSION: With stitches maintained and without intervention, the acute rejection is similar in rat corneal transplantation and human corneal transplantation. It is explained in the experiment that 8 stitches in corneal transplantation in which the rat implant tissue of 3.5 mm matches graft foreman of 3.0 mm can reduce the complications of rat corneal transplantation and becomes a satisfactory animal model of corneal transplantation.Skillful microscopic operation techniques, satisfactory operation instruments and adequate amplilfied pupil can reduce to the most extent the postoperation complications in rats.
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In order to investigate the ipsilateral lymphadenectomy for inhibiting rejection in rat corneal transplantation, corneal allogenic transplantation models were established in rats. Eighteen female Wister rats were used as donors, and 36 Sprague Dawley rats as recipients. After penetrating corneal transplantation, recipients were randomly divided into 3 groups: group A (control group);group B, the ipsilateral lymphadenectomy group; group C, the bilateral lymphadenectomy group.Among 12 rats in each group, the corneas of 2 rats in each group were used for pathological study at day 14 after the transplantation, and the remaining 10 rats were used for studying corneal rejection by a slit lamp. The time points when allograft rejection occurred were recorded and mean survival time (MST) was compared. The results showed that MST in groups B and C was 46.30±9.464 days and 44.43 ± 7. 604 days, respectively, which was significantly prolonged as compared with that in group A (10.71±1. 567 days, P＜0.01). There was no significant difference in MST between groups B and C (P＞0.05). Itwas concluded that both bilateral and ipsilateral lymphadenectomy therapies could effectively inhibit the corneal allograft rejection. Ipsilateral lymphadenectomy is a less complex surgical procedure and is just as effective in preventing rejection.
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The expression of VEGF-C and molecular mechanisms of lymphangiogenesis in rat cornea after alkali injury was studied. The rat alkali injured corneal models were made. Under electron microscopy, the lymphatic vessels in the rat injured corneas were examined. The expression of VEGF-C proteins was detected by using immunohistochemical assay at day 1, 3, 5, 7, 9 after injury. The expression levels of VEGF-C mRNA were quantified with reverse transcription polymerase chain reaction (RT-PCR). The results showed that the lymphatic vessels were found in the injured rat corneas 14 days after the injury. The VEGF-C protein was detectable 3 days after injury, reached the peak 5 days after injury, and gradually decreased. In the control group, no VEGF-C proteins were detected. The VEGF-C mRNA was minimally detected in the normal rat corneas, but it was highly expressed 5 days after the injury. The difference was statistically significant. It was concluded that VEGF-C might be one of the most important relevant factors in corneal lymphangiogenesis after alkali injury.
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Animais , Ratos , Biomarcadores , Queimaduras Químicas , Metabolismo , Córnea , Metabolismo , Lesões da Córnea , Queimaduras Oculares , Metabolismo , RNA Mensageiro , Genética , Distribuição Aleatória , Ratos Sprague-Dawley , Fator C de Crescimento do Endotélio Vascular , GenéticaRESUMO
To investigate whether the TGF-beta 1 plasmid DNA carried by lipofectamine could be introduced into cultured rabbit corneal epithelial cells, specific expression of the plasmid pMAM TGF-beta 1 in the cultured corneal epithelial cells was studied. Two days after 12 h of transfection of pMAMT-GF-beta 1 mediated by lipofectamine into the cultured corneal epithelial cells, the TGF-beta 1 protein expression specific for pMAMTGF-beta 1 in the cells was detected by means of immunohistochemical staining and the positive rate was 23.37%. The results suggested that foreign plasmid DNA could be effectively delivered into cultured rabbit corneal epithelial cells by means of lipofectamine, and this will provide a promising method of studying TGF-beta 1 on the mechanism of physiology and pathology concerned with corneal epithelial cells.
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Animais , Coelhos , Divisão Celular , Células Cultivadas , Epitélio Corneano , Biologia Celular , Metabolismo , Técnicas de Transferência de Genes , Vetores Genéticos , Lipídeos , Farmacologia , Plasmídeos , Genética , Fator de Crescimento Transformador beta , Genética , Fator de Crescimento Transformador beta1RESUMO
The method of gene transfer into corneal endothelium was investigated to provide a foundation for the study of TGF-beta 1 gene transfer to inhibit corneal graft rejection. Two days after direct injection of pMAM TGF-beta 1 mediated by liposome into the anterior chamber of rabbits, one half of corneas were made into paraffin slides and the endothelial layer was carefully torn from the other half to make a single layer slide of endothelia. By means of immunohistochemical technique, the plasmid pMAM TGF-beta 1 expression product TGF-beta 1 in the endothelia was detected. Specific TGF-beta 1 expression was positive in the endothelia on both the paraffin slide and the single layer slide. The results showed that by direct injection into the anterior chamber, foreign plasmid DNA could be transferred into the endothelia and its expression was obtained. This may provide a foundation for further study on TGF-beta 1 participating in local induction of corneal immune tolerance.