1.Construction and activity identification of recombinant retroviral vector expressing bone morphogenetic protein 2 gene
Qing LIAO ; Ying TANG ; Renjie CHI ; Xiaochun CHEN ; Jingyu GUAN ; Youwen DUAN
Chinese Journal of Tissue Engineering Research 2010;14(7):1227-1230
BACKGROUND: Bone morphogenetic protein (BMP) is a protein possesses potential activity, which can increase and enhance its activity when the bone issues are damaged, so it can be used to repair the bone defects when combined with carrier. However,there are few reports concerning it as gene therapy.OBJECTIVE: To construct recombinant retroviral vector expressing human BMP2 gene and to discuss its biological function in ostecblasts.METHODS: BMP2-specific primers were designed and synthesized according gene sequence of human BMP2 gene in Genbank, then BMP2 gena was amplified by Hifi PCR, which was recombined with cloning vector pDNR-CMV homologousiy into pDNR-CMV-BMP2 plasmid identified by BMP2 PCR and enzyme digestion of Sail and EcoRI as well as gene sequencing; recombinant plasmid pDNR-CMV-BMP2 and retroviral plasmid pLP-LNCX were recombined homologously in IoxP sites into pLP-LNCX-BMP2 plasmid transferred into packing cell line PT67 and the supernatant was collected to assay viral titre. Human osteoblast was infected with retrovirus, then the growth of cells were observed by MTT, and the expression of BMP2 protein was detected by Western blotting at 48 hours transfectionRESULTS AND CONCLUSION: Digestion, BMP2 PCR and gene sequencing of pDNR-CMV-BMP2 were coincided with expected. After transfected with plasmid pLP-LNCX-BMP2, PT67 cells could be screened with G418 only to get stably integrated in BMP2, of whose supemanant viral titra amounted to 5×10~8 pfu/mL. MTT assay showed that there had no evident difference in cellular inhibition between the normal and retrovirus groups at 72 hours after transfection (P > 0.05); Western blotting showed that the BMP2 was strong expressed at 48 hours after transfection. It demonstrated that BMP2 gene was successful cloned and its retrovirus vector was constructed.
2.Three-dimensional CT angiography study on the relations between the vertebral artery and atlantoaxial joint.
Shao-yin DUAN ; Shao-mao LÜ ; Feng YE ; Qing-chi LIN ; Liao-bin CHEN
Chinese Medical Journal 2009;122(8):917-920
BACKGROUNDThe vertebral artery (VA) and atlantoaxial joint (AAJ), with complicated structures, are located in the depths of the head-neck boundary area, the regional anatomy of which cannot be shown globally and directly. This study aims to evaluate three-dimensional CT angiography (3DCTA) in displaying the AAJ, atlantoaxial segment of the vertebral artery (ASVA) and the identification of their interrelations.
METHODSSixty-eight subjects without pathology of the ASVA and AAJ were selected from head-neck CTA examination. All the 3D images were formed with volume rendering (VR) together with techniques of separating, fusing, opacifying and false-coloring (SFOF). On the 3D images, the ASVA and AAJ were observed, and their interrelations were measured.
RESULTSAll the 3DCTA images were of high quality and up to our requirements. They could clearly and directly show the ASVA, ascending along the AAJ. There were 5 curves in the course of the ASVA, of which 2 curves were away from the atlantoaxial joint, one in the 2nd curve of 0.0 mm - 5.4 mm, the other in the 4th of 2.6 mm - 9.2 mm. There was no significant difference in the measurements between left and right (P > 0.05). The curved parts of the ASVA slightly expanded, with the biggest diameter of 5.6 mm in the 4th curve. Statistical comparison shows that the left ASVA is larger than the right (P < 0.05). Variations of the ASVA were found in 8 cases and of the AAJ in 12.
CONCLUSIONS3DCTA can globally and directly demonstrate the structures of the AAJ, ASVA and their interrelations. The 3D imaging data make up and enrich the research contents of regional anatomy and lay the foundation for related study and applications.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Atlanto-Axial Joint ; diagnostic imaging ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Vertebral Artery ; diagnostic imaging ; Young Adult
3.Effect of estrogen replacement therapy in the early phase on atherosclerosis and PAI-1 in ovariectomized cholesterol-fed rabbit.
Hai-Yun DONG ; Xiang-Ping LI ; Ya-Jun LIAN ; Qing-Chi LIAO
Journal of Central South University(Medical Sciences) 2007;32(4):626-630
OBJECTIVE:
To investigate the effect of estrogen replacement therapy (ERT) in the early phase on the atherosclerosis and the level of plasminogen activator inhibitor-1(PAI-1).
METHODS:
Twenty-eight rabbits were randomly assigned to 4 groups: Group A, sham operation (n=7); Group B, ovariectomized without estradiol (n=7); Group C, ovariectomized with low-dose estradiol (n=7); and Group D, ovariectomized with high-dose estradiol (n=7). All rabbits were given 1% cholesterol diet for 12 weeks. Levels of blood lipid, estradiol, and PAI-1 were measured before the operation and at the end of the 4th and 12th weeks. Twelve weeks later, we took the aortas for pathological analysis and calculated the areas of atherosclerotic plaque.
RESULTS:
After 12 weeks, the estradiol level of Group B was significantly lower than that of Group A, and that of Group D was obviously higher than Group A. There was no significant difference between Group C and A. The concentrations of total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) in Group B significantly increased compared with Group A (P<0.01). The levels of TC and LDL-C of Group C and D were significantly lower than those of Group A. Whereas the concentrations of triglyceride (TG) and high density lipoprotein cholesterol (HDL-C) in Group B were lower than those of Groups A, C and D (P<0.01). In contrast to Groups A, C and D, the level of PAI-1 was significantly higher in the Group B (P<0.01), without significant differences among Groups A, C and D. The area of atherosclerotic lesion of aorta in Group B was significantly bigger than that of Group A, C and D. The areas of aortic atherosclerotic plaque in Group C and D were obviously smaller than those of Group A (P<0.01).
CONCLUSION
Transdermal estrogen replacement therapy in the early phase can improve the metabolism of the serum lipids, reduce the level of PAI-1, and probably provide the protective effect on the atheroma formation.
Administration, Cutaneous
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Animals
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Atherosclerosis
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blood
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drug therapy
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pathology
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Cholesterol
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blood
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Estradiol
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administration & dosage
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Estrogen Replacement Therapy
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Female
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Ovariectomy
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Plasminogen Activator Inhibitor 1
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blood
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Rabbits
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Triglycerides
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blood
4.Preservation of the pituitary stalk and the gland in transsphenoidal microsurgery for pituitary adenomas.
Haoyu LI ; Xianrui YUAN ; Yiwei LIAO ; Yuanyang XIE ; Chi ZHANG ; Juan LI ; Jun SU ; Xiangyu WANG ; Xiaoyu CHEN ; Qing LIU
Journal of Central South University(Medical Sciences) 2014;39(2):136-141
OBJECTIVE:
To improve the surgical outcome of pituitary adenomas by identifying and preserving the pituitary stalk and the gland during surgery.
METHODS:
From October 2010 to September 2012, the author from the Department of Neurosurgery of Xiangya Hospital, Central South University operated on 51 patients with pituitary adenoma. During the operations, we carefully identified the normal adenohypophysis, pituitary stalk, neurohypophysis and the abnormal tissues either by direct observation or by medical images, aiming to excise the tumor thoroughly, protect the pituitary function and reduce the postoperative complications.
RESULTS:
Totally 37 patients (72.5%, 37/51) had total resection of the tumor, 12 (23.5%, 12/51) had subtotal tumor resection and the other 2 had major removal. The gland and the pituitary stalk were well identified and reserved. Detection of hormone content proved that the operation had little effect on the free triiodothyronine (FT3) and adrenocorticotropic hormone (ACTH), while for free tetraiodothyronine (FT4) and thyroid stimulating hormone (TSH) and postoperative followup significant alleviation was found. There was no significant fluctuation for the testosterone in the men preoperatively and postoperatively (all the above results were obtained without hormone replacement therapy). The main postoperative complications were as follows: temporary diabetes insipidus in 5 patients (9.8%, 5/51); electrolyte disorder (the appearance of hyponatremia) in 17 (33.3%, 17/51); and cerebrospinal fluid rhinorrhea and postoperative intracranial infection in 1 (2%, 1/51). No one died during the perioperation period.
CONCLUSION
Microscopic transsphenoidal surgery is effective for pituitary adenomas including tumors violating the cavernous sinus. Accurate identification of the pituitary stalk, the gland and the abnormal tissue during the microscopic transsphenoidal operation plays a critical role in preserving the pituitary function and promoting postoperative rehabilitation.
Adenoma
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surgery
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Humans
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Male
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Microsurgery
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Neurosurgical Procedures
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methods
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Pituitary Gland
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surgery
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Pituitary Hormones
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blood
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Pituitary Neoplasms
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surgery
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Postoperative Complications
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Treatment Outcome
5.Comparative analysis of the management practice of including designated retail pharmacies in the outpatient pooling of basic medical insurance
Qing-Qiang LEI ; Tian-Chi CHEN ; Bo-Wei LIAO ; Zi-Le YAN ; Hong XU ; Wen CHEN ; Min HU
Chinese Journal of Health Policy 2023;16(12):26-32
Objective:To summarize and analyze the practice of the local practice on covering drugstore bills by pooled funds of basic medical insurance,and provide a reference for improving relevant policies.Methods:The medical insurance policies from various provinces,municipalities,autonomous regions,and coordination areas were systematically retrieved.ROST CM6 software was applied to analyze the high-frequency words and semantic network of the policy text,and combined with the interview and field investigation resultss,the key dimensions of the policy practice were identified and summarized.Then the regional differences,existing problems,and their causes were analyzed to put forward policy recommendations.Result:The selection of pharmacies covered in the payment system with outpatient expenses reimbursed by the pooled fund,the formulation of drug reimbursement list,the design of benefit plans,the management of drug prices and payments,and the supervision of hospital outflow prescriptions were five key dimensions of policy practice.There were significant differences in practice among different regions,and the problems mainly included the overall arrangement of covering pharmacies in the payment system,the mechanism of drug prices in pharmacies,and the coordination with other medical insurance policies.Conclusion:To improve the convenience of buying drugs for the insured,it is necessary to make full use of the advantages of pharmacies to meet the demand for outpatient medicine,promote the transparency of drug prices in pharmacies,coordinate the relevant medical insurance policies,strengthen the collaborative management between the healthcare security administration and relevant departments such as the health commission and the medical products administration,analyze and evaluate the potential effects of policy measures,and adjust policy measures promptly according to local conditions.