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 investigate the transfection and expression of recombinant plasmid human vascular endothelial growth factor 165/pcDNA3. 1 （hVEGF165/pcDNA3. 1） in myocardial cells, and to build foundation for gene therapy and cell therapy of coronary artery disease （CAD）. Methods Myocardial cells were cultured in vitro and transfected by hVEGF165/pcDNA3.1 with liposome; then transient expressed protein was detected by reverse transcriptase-polymerase chain reaction （RT-PCR）, immunochemistry and Western blotting. Results A strap as hVEGF165 was obtained by RT-PCR, the protein of hVEGF165 was found in myocardial cells by immunochemistry and in supernatant by Western blotting. Conclusion The recombinant plasmid hVEGFI65/pcDNA3. 1 can be expressed in myocardial cells, and may be used in studying CAD by gene therapy and cell transplantation.
【 Abstract 】 Objective To study the mRNA expression of BC047440 gene in multiplicate malignant tumor tissues and the corresponding adjacent tissues, and to investigate its roles in the carcinogenesis and development of malignant tumors. Methods Forty-eight cases of malignant tumor tissues and their adjacent non-cancerous tissues were examined. The mRNA expression of BC047440 gene in those tissues of liver cancer, cholangiocarcinoma, gastric cancer, carcinoma of large intestine, glioma, and breast cancer were measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results ① The mRNA expressions of BC047440 gene in liver cancer, gastric cancer, cholangiocarcinoma and carcinoma of large intestine were significantly higher than those in their adjacent non-cancerous tissues (Plt;0.05 or 0.01). BC047440 gene were highly expressed in both glioma and its adjacent tissues (Pgt;0.05), and poorly expressed in both breast cancer and its adjacent tissues (Pgt;0.05). ② There were close relationships between BC047440 gene expression and clinicopathologic findings of liver cancer, including tumor size and portal vein invasion (Plt;0.05). ③ There were also close relationships between BC047440 gene expression and different clinical stages in alimentary canal cancers (Plt;0.05). Conclusion The over expression of BC047440 gene may be related with the growth, infiltration and metastasis of some malignant tumors, including liver cancer, cholangiocarcinoma, gastric cancer, carcinoma of large intestine and glioma.
Objective To investigate the expression of ADAM9 in breast cancer and its clinical significance. Methods The expressions of ADAM9 in normal breast tissues and breast cancer tissues were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry, and whose relationship with clinicopathologic features was analyzed. Results The expression of ADAM9 mRNA increased in the breast cancer tissues, but which was not detected in the normal breast tissues. The expression of ADAM9 protein in the breast cancer tissues was significantly higher than that in the normal breast tissues (Plt;0.05), and which in the metastatic lymph nodes was significantly higher than that in the negative lymph nodes or corresponding primary lesions (Plt;0.05). The expression of ADAM9 in the breast cancer tissues was correlated with the lymph node metastasis and histological grade (Plt;0.05). Conclusion ADAM9 is overexpressed in the breast cancer tissues, which might involve in the pathological progression of breast cancer.
We have devised a highly sensitive, specific, and quantitative assay for multidrug resistance (mdr1) mRNA expression based on the reverse transcription-polymerase chain reaction (RT-PCR). mdr1 mRNA levels were detected in 30 human primary hepatocellular carcinoma (PHC) tissue and adjacent liver tissue. Five of the patients had received chemotherapy before hepatectomy. The results show that the level of expression of mdr1 gene is higher in tumor tissue than in adjacent liver tissue. mdr1 gene is overexpressed in PHC after chemotherapy. Furthermore, mdr1 gene expression in the treated tumor adjacent liver tissue is higher than that in untreated tumor adjacent liver tissue. Our results indicated that overexpression of mdr1 gene may be responsible for the intrinsic and acquired drug resistance of PHC.
ObjectiveTo investigate the clinical significance of CK20 mRNA expression in blood of patients with colorectal cancer. MethodsThe expressions of CK20 mRNA in blood of twenty healthy volunteers, ten patients with colorectal polyp and sixtyone patients with colorectal cancer were detected by RT-PCR. ResultsThe positive rate of CK20 mRNA in peripheral venous blood and portal venous blood of patients with colorectal cancer were 41.0%(25/61) and 45.9%(28/61), which was not significantly different (Pgt;0.05). The expression of CK20 mRNA in patients with colorectal cancer was associated with clinical TNM stage of tumor, local lymph node metastasis, distance metastasis, and the depth of invasion (Plt;0.05). No expression of CK20 mRNA was detected in blood of twenty healthy volunteer’s and ten patients with colorectal polyp. ConclusionCK20 is a specific marker for detecting blood micrometastasis of colorectal cancer. The expression of CK20 mRNA in blood of patients with colorectal cancer is related with TNM stage, invasion, and metastasis of colorectal cancer.
OBJECTIVE: To provide anatomical bases for dorso-ulnar aspect of mid-hand reverse flap. METHODS: After red latex was infused into the arteries of 40 sides of adult cadava upper limbs, the origin, course, branches, distribution and distal anastomosis on the dorsal carpal branch of ulnar arteries were observed. And the mid-hand flap transfer was used to repair two cases of soft tissue defect (ranged 4.5-5.0 cm x 2.0-3.5 cm on ring and little fingers). RESULTS: The dorsal carpal branch begins with ulnar artery (3.9 +/- 1.2) cm above the pisiform with diameter of (1.3 +/- 0.2) mm, and branches off into ascending and descending branches. The descending one is the continuing of dorsal branch, it crosses the ulnar edge of the fifth metecarpal bone and anastomizes with the digital artery of little finger or hypothenar branch of deep palmar (accounted for 70%). While the other ascending branch with the former two branches formed anastomosis accounts for 30%. The two cases got healed in one-stage. The function of fingers recovered after 3-4 month follow-up. CONCLUSION: The reverse flap of dorso-ulnar aspect of mid-hand is available to repair the soft tissue defect on dorsum of hand with neighbor finger.
To introduce a new technique for vascular pedicle elongation in the anterolateral thigh island flap transplantation and evaluate the outcome of this technique in the clinical application. Methods From January 2003 to January 2006, 6 patients (5 males, 1 female; age, 1849 years) were admitted for surgical operation because of the soft tissue defect around the knee joint. The soft tissue defect after the injury was found in 3 patients, the defect after the removal of the softtissue tumor in 1, and the defect after the prosthetic replacement in the knee joint in 2. The soft tissue defects ranged in size of 8 cm×4 cm to 15 cm ×6 cm. When the anterolateral island flap of the thigh underwent the reverse transplantation, the ascending branch of the lateral circumflex femoral artery was used as a nutrient vessel for the flap, and the descending branch of the lateral circumflex femoralartery was separated to the distal part. The main trunk of the lateral circumflex femoral artery was ligated at the point that was proximal to the furcation ofthe ascending and decending branches so that the vessel pedicle of the flap could be lengthened and then the defect was repaired.The flaps ranged in size of 10cm×6 cm to 18 cm×8 cm Results All the flaps were successfullytransferred in the 6 patients. The lengthened pedicle ranged in length from 8 to 12 cm, with an average of 10 cm. There was no vascular crisis after operation. All the transferred flaps survived, with a color and texture similar to those in the recipient site. The postoperative followup for 6-18 months revealed that the motion range of the knees was satisfactory. Conclusion The vascular pedicle elongation technique can enlarge the application scope of the anterolateral thigh island flap and the survival rate of the flap is not influenced by any factor.
Objective To investigate the operative procedure and the cl inical results of reverse lateral tarsal artery flap in treating forefoot skin and soft tissue defect. Methods From August 2007 to April 2009, 11 patients with forefoot skin and soft tissue defect were treated with reverse lateral tarsal artery flaps, including 7 males and 4 females aged from 16 to 60 years(36 years on average). Of 11 cases, defects were caused by crash in 5 cases, by grind contusion in 3 cases and the course disease was 4-12 hours; by tumor extended resection in 3 cases and the disease course was 3-12 months. There were 5 wounds on the dorsum of first metatarsophalangeal joint, 2 on the dorsum of the first toes, and 4 on the dorsum of distal part of metatarsal bones. The area of defect ranged from 4 cm × 2 cm to 6 cm × 5 cm. There were 6 cases of tendon exposure, 4 cases of tendon defect with bone exposure, and 1 case of tendon defect with open dislocation of metatarsophalangeal joint. The flap was designed with dorsal artery of foot as its pedicle. The plantar perforating branch was designed as its rotating point. And the flaps were transferred retrogradely to repair the forefoot wounds. The flap area ranged from 4.5 cm × 2.5 cm to 6.5 cm × 4.5 cm. The lateral dorsal nerve of foot was anastomosed with the nerve in wound area in 7 cases. Donor site was covered by full thickness skin graft. Results Partial necrosis occurred and was cured by dressing change, followed by skin graft in 2 cases. The flaps survived and primary heal ing was achieved in the other 9 cases. All the skin grafts of donor site survived and primary heal ing wasachieved after operation. All the patients were followed up for 6 months to 2 years, averaged 13 months. The texture and color of the flap were similar to skin at the recipient site. All patients returned to normal in walking and running and no ulceration occurred. The two point discrimination was 5-12 mm 6 months after operation in 7 patients who received nerve anastomosis, while only protective sensation recovered partly in the other 4 patients whose cutaneous nerve were not anastomosed. Conclusion Reverse lateral tarsal artery flap has the perfect shape and its blood vessel is constant. The blood pedicle is thick and long enough when transferred retrogradely. The flap is a good choice in the treatment of forefoot skin and soft tissue defect.
OBJECTIVE: To investigate the effect of electroacupuncture on mRNA expression of NGF and IGF-1 in injured nerve. METHODS: Sciatic nerve injury model was established by transection of right side sciatic nerve in 90 male SD rats, which were randomly divided into two groups. The experimental group was treated with electroacupuncture, no treatment in the control group. The distal part of the injured nerve was harvested after 1, 2, 4, 6 and 10 weeks of operation and stored in the liquid nitrogen. The total RNA was extracted by the TRIzol reagent. Reverse transcriptase-polymerase chain reaction(RT-PCR) was used to detected the mRNA expression of NGF and IGF-1. RESULTS: The mRNA expression of NGF in the experimental group was increased quickly from the second week, and reached to highest level in the fourth week. It was much higher than that of the control group (P lt; 0.05). Then it began to decline in following time and approximately reached to the level of the first week after 10 weeks of operation. The mRNA expression of IGF-1 in the experimental group was remarkably increased in the second and fourth week, and which was much higher than that of the control group respectively(P lt; 0.05). Although the mRNA expression of IGF-1 after 10 weeks of operation in the experimental group was higher than that of the control group, but there was no significant difference between the two groups(P gt; 0.05). There was linear correlation in the fourth week between mRNA expression of NGF and IGF-1 in the experimental group. CONCLUSION: The mRNA expression of NGF and IGF-1 can be elevated in injured nerve at early stage interfered with electroacupuncture.