1.Effect of basic fibroblast growth factor gene transfection on bio-behavior of rat extraocular muscle satellite cells
Ligui ZHANG ; Chuanfu WANG ; Xiaohui HUANG
Chinese Journal of Tissue Engineering Research 2007;11(33):6706-6711
BACKGROUND: Experiments have demonstrated that exogenous basic fibroblast growth factor (bFGF) promotes the growth and survival of skeletal muscle satellite cells, and endogenous bFGF also has obvious effect on muscular repair.But whether bFGF gene transfection has the same effect on extraocular muscle satellite cells is unclear.OBJECTIVE: The goal of this study was to investigate the effect of bFGF gene transfection on the bio-behavior of rat extraocular muscle satellite cells.DESIGN: Single sample observation.SETTING: Department of Ophthalmology, Qingdao University Medical College.MATERIALS: Primary rat extraocular muscle satellite cells were cultured (purity > 90%). Human PEGFP-N3-bFGF eukaryotic expression vector was successfully constructed (reported in other papers).METHODS: This study was carried out in the Central Laboratory of Qingdao University Medical College between September 2005 and December 2006. Experimental intervention and grouping: During the experiment, 3 groups were divided: Experimental group, in which, recombinant PEGFP-N3-bFGF was used for transfection; Control group A, in which, empty-load PEGFP-N3 was used for transfection; Control group B: in which, only F 10 medium (0.1 volume fraction of fetal bovine serum)was used for transfection. Recombinant PEGFP-N3-bFGF plasmid was used to transfect rat extraocular muscle satellite cells cultured in vitro by liposome-mediated transgenic technology. Experimental evaluation: bFGF expression was observed by immubohistochemical method; Transfection efficiency was detected with fluorescence microscope; The protein secretion of bFGF of satellite cells in each group was detected by SBC-ELISA method on the 1st, 3rd, 5th, 9th, 11th, 13th, 15th, 21st and 28th days of culture; The proliferation of rat extraocular muscle satellite cells in each group was detected by methyl thiazolyl tetrazolium(MTT) method; Creatine kinase (CK) activity of cells in each group was determined according to the A value of some standard whose concentration was known.MAIN OUTCOME MEASURES: ① Transfection efficiency of bFGF gene to rat extraocular muscle satellite cells cultured in vitro and bFGF gene expression after transfection; ②bFGF protein secretion after transfection, and effect of bFGF gene transfection on the growth and proliferation of extraocular muscle satellite cells.RESULTS: ① Transfection efficiency of bFGF gene to rat extraocular muscle satellite cells cultured in vitrowas (42.8±1.2)%. ② Immunohistochemical detection showed positive reaction. ②Transfected cells could secrete bFGF protein,which was the highest on the 5th day (662.935 ng/L). ④ The proliferation activity of transfected cells was obviously enhanced, and its A value was significantly higher than that of control group at the same time point (P < 0.05). ⑤CK value was higher than that of the control group from the 5th day after transfection to the end (P < 0.01).CONCLUSION: bFGF gene transfering into rat extraocular muscle satellite cells can make extraocular muscle satellite cells to secrete bFGF, promote cell proliferation, survival and differentiation.
2.Expression of the transfected basic fibroblast growth factor gene in myoblasts and regulatory system
Ligui ZHANG ; Hongyun WANG ; Leilei QIN ; Xiaohui HUANG ; Chuanfu WANG
Chinese Journal of Tissue Engineering Research 2010;14(20):3780-3786
BACKGROUND:Transgenosis of basic fibroblast growth factor (bFGF) gene has been successfully performed into the muscle satellite cells of rat extraocular muscles in the previous study of the research group, proving that bFGF could express in the myoblasts of extraocular muscles, also promote cell proliferation and differentiation.OBJECTIVE: To further investigate the methods for regulating the expression of the bFGF in myoblasts following transfection. METHODS: Target gene bFGF was connected with inducing expression vector pcDNA4/T0/myc-His?A, followed by masculine clone sequencing identified by colony PCR and enzyme digestion, EcoR I and Hind III restriction enzyme digestion, as well as Xho I single enzyme verification. C2C12 myoblasts antibiotics sensitivity was screened and finally defined. By use of lipofection transfection technology, cell lines where C2C12 stably expressed pcDNA6/TR were estabolishd and then identified by Western blot. The pcDNA4/TO/myc-His?A-bFGF was transfected into pcDNA6/TR- C2C12 cells. The bFGF expression and secretion in C2C12 cells following tetracycline-induced pcDNA4/TO/myc-His?A-bFGF transfection were determined by immunofluorescence and Western blot, the controls were established.RESULTS AND CONCLUSION: ① The conjunction between the bFGF and inducing expression vector pcDNA4/TO/myc-His?A was proved successfully by sequencing comparison, double digestion and single digestion. ②The minimal lethal concentration of blasticidin to C2C12 cells was 10 mg/L, while that of zeocin was 750 mg/L. ③ The pcDNA6/TR-C2C12 cell lines were established correctly. ④ The myoblasts treated by tetracycline and transfected with pcDNA4/TO/myc-His?A-bFGF were positive for gene expression, those untreated exhibited a negativity; bFGF protein could be produced in myoblasts treated by tetracycline and transfected with pcDNA4/TO/myc-His?A-bFGF, the production reached a peak at 24 hours, while those untreated can not produce bFGF protein. Results suggest that the bFGF expression in the myoblasts can be controlled by tetracycline inhibition and regulatory systems.
3.The comparison research of intrathecal injection of oxygen and hyperbaric oxygen on the rehabilitation of patients with craniocerebral injury
Ligui GAO ; Huifang WANG ; Wei SHI ; Ruizhi WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2004;0(05):-
Objective To investigate the role of intrathecal i njection of oxygen in the treatment of patients with craniocerebral injury throu gh the comparison research of intrathecal injection of oxygen and hyperbaric oxy gen on the rehabilitation of patients with craniocerebral injury. Methods A total of 150 patients were divided into three gro ups: the group of intrathecal injection of oxygen, the group treated with hyperb aric oxygen and the group of intrathecal injection of air. CSF was applied via l umbar puncture. The rehabilitative function of patients was observed and the lev el of superoxide dismutase (SOD), malondialedhyde (MDA), and nitrogen monoxide ( NO) in CSF were measured. Results The neurological function of the patients treated w ith oxygen injected intrathecally intrathecal injection of oxygen was improved w as more significanty than that of patients treated with hyperbaric oxygen; the i ncrease of SOD, NO and the decrease of NO were more significant than that in the group treated with hyperbaric oxygen. Conclusion Intrathecal injection of oxygen can improve the rehabilitative function more significantly than the hyperbaric oxygen therapy in the treatment of craniocerebral injury.
4.Diagnosis and treatment of renal tuberculosis
Ligui MIN ; Bin WEN ; Yinggang WANG ; Hongliang JIA
Chinese Journal of Urology 2010;31(11):761-763
Objective To discuss the diagnosis and treatment of renal tuberculosis.Methods A retrospective study was made on 96 cases.Results Frequency (51.8%), urgency (37.2%),odynuria (33.4%),lumbodynia (41.0%),and hematuria(48.1%)were the most common symptoms. The diagnostic accuracy of IVU, B-type ultrasonography, CT and biopsy of mucous membrane of urinary bladder were 69.1%,12.5 %,37.5% and 33.3% respectively. 96 cases were given medicine (INH+RFP+PZA or PIA for 6-8 months).38 cases(39.6%)have been cured, while symptoms of 43 cases (44.8%) have been improved. Operation was performed on 15 cases that were ineffective treated by chemical therapy. Conclusions Urine routine, IVP, cystoscopy+biopsy of mucous membrane of urinary bladder provide important information for the diagnosis of renal tuberculosis. INH, REP and EMB or PIA combination therapy yields satisfactory outcome for early cases.
5.Pathogenic mechanisms of influenza virus and Staphylococcus aureus co-infection:research progress
Yuan LIANG ; Ligui WANG ; Shaofu QIU ; Leili JIA ; Hongbin SONG
Military Medical Sciences 2017;41(5):406-409
Studies have shown that co-infection of influenza viruses with bacteria is an important cause of high mortality during the epidemic of influenza.There are at least 12 species of bacteria that have been reported to be able to co-infect with influenza.Among those species,co-infection with Staphylococcus aureus is not only the most common but also the most lethal.However,the pathogenesis of high mortality from co-infection with influenza virus/S.aureus remains elusive.In addition,co-infection of influenza virus/S.aureus can induce severe pneumonia.There is new evidence that influenza virus can reduce the host′s tolerance to pathogenic or inflammatory injury,and the two pathogens can also synergistically aggravate toxic effects on the host.Here,we review the mechanisms of severe mortality of influenza infection associated with S.aureus co-infections in order to contribute to prevention and control of influenza in the future.
6.Application of Excel Office Software in the Management of Clinical Teaching
Liangjun WU ; Mingzhao XIAO ; Ligui WANG ; Lingquan KONG ; Zhengzhao LIU
Chinese Journal of Medical Education Research 2003;0(03):-
With the development of computer technique, Excel office software is widely applied in the management of clinical teaching. According to the problems existing in the clinical teaching management of our college, Excel electronic software is accepted to reform the present management measures. Its designation, essential archives establishment, method, result, analysis and discussion are introduced definitely in this paper.
7.Comparison of retroperitoneal mini-incision and laparoscopic methods in treatment of varicocele
Yonghu SHEN ; Ligui MIN ; Hesuti MA ; Jianjun ZHANG ; Yun SUN ; Bo XIE ; Yong CHU ; Zhuqian WANG ; Qun WU
International Journal of Surgery 2009;36(12):805-807
Objective To evaluate the best mean for the treatment of varicocele. Methods The clini-cal effects of the high ligation of spermatic vein under the small incisal opening of the poritneum meta abdom-inal membrane and the ligation of spermatic vein under the laparoscope for the varicocele were retrospectively reviewed. The operating time, the postoperative days and the cost of hospitalization were compared of the two methods. Results Comparing with the group of operation and the group of laparoscope. Among the 158 vat-icocele patients with unilateral or bilateral varicocele, there was not statistical difference in the operating time between the two methods (P > 0. 05), whi]e there was significant difference in the postoperative days and the cost of hospitalization(P <0. 05). Conclusions When the bilateral varicoceles were simultaneously ligated, the high ligation of spermatic vein under the laparoscope was the best selection; if the unilateral varicocele was highly ligated or the patients were unconditional or the patients were not fit for laparoscopic operation, the high ligation of spermatic vein under the small incisal opening of peritoneum was the best operating method.
8.RXPERIMENTAL AND CLINICAL STUDY OF CREATINE KINASE BB ACTIVITY FOR TH E DIAGNOSIS ON BRAIN DAMAGE
Wei SHI ; Fangru WANG ; Ren ZHOU ; Ligui GAO ; Jianqiang QU ; Jianjun SUN ; Shouping GONG ; Chongxiao LIU ; Le ZHOU
Journal of Pharmaceutical Analysis 2001;13(2):113-115
Objective To study the differential diagnosis o n cerebral concussion and mild cerebral contusion value of the brain type creati n e kinase isoenzyme(CK-BB),and evaluate the seriousness of brain damage and prog nosis of the patients with acute head injury.Methods Chromatographic separating and fluorometric quant ifying technique was used to detect the CK-BB activity in the cerebrospinal flu id(CSF) of 117 patients with acute head injury and 12 patients with increased in tracranial pressure and 20 normal people.Results The CSF-CK-BB activity of the patients with acu te head injury was remarkably higher than that of the normal people and the CSF -CK-BB activity increased with the seriousness of brain damage.There was a clo se relationship between CSF-CK-BB activity and prognosis,and higher activity o f CSF-CK-BB indicated poor prognosis.Conclusion CSF-CK -BB activity could be used as a new index to diagnose brain damage and evaluate the seriousness of brain damage and prognosis.
9.Correlation analysis between vitamin A, D and E levels with the altitude, seasonal variation and other factors in children with 0-6 years old in Tibetan Plateau of Ganzi Prefecture
Ping HUANG ; Xinmei LIN ; Quansheng WANG ; Gang KE ; Zhengrong WEI ; Duping GUO ; Xueguo BA ; Yongxiao YUAN ; Qun WANG ; Hongyan DING ; Maoyun HE ; Ligui XIAN ; Liping ZHANG ; Yufei WANG ; Lianhui YU ; Kehong KUANG ; Qingping LUO ; Zhu CHEN ; Hongmei LI ; Jing LUO ; Yilan XU
Chinese Journal of Applied Clinical Pediatrics 2021;36(22):1736-1741
Objective:To detect serum levels of vitamin A (Vit A), vitamin D(Vit D)25-hydroxy vitamin D[25-(OH)D] and vitamin E(Vit E) in children aged 0-6 years in Tibetan Plateau of Garzi Prefecture, thus providing references for physical examinations and prevention of 4 key diseases (rickets, malnutrition anemia, pneumonia and diarrhea) in children in plateau areas by relevant government departments.Methods:A total of 2 122 children who participated in physical examination in 12 townships of Xiangcheng County and 14 townships of Daocheng County, Garzi Tibetan Autonomous Prefecture, Sichuan Province from April 2017 to April 2019 with 0-6 years old were recruited for surveying physical measurements and collection of venous blood.Serum Vit A and Vit E levels were detected by high performance liquid chromatography.Serum levels of 25-(OH)D were detected by high performance liquid chromatography tandem mass spectrometry.The relationship between Vit A, Vit E and 25-(OH)D levels with the gender, age, seasonal change and altitude was analyzed.Results:The serum Vit A level, subclinical Vit A deficiency rate and marginal vitamin A deficiency rate were(1.05±0.27) μmol/L, 8.15%(173/2 122 cases) and 45.99%(976/2 122 cases), respectively in 2 122 children with 0-6 years old.There were significant differences in the serum Vit A level, the subclinical Vit A deficiency rate and the marginal vitamin A deficiency rate in children with different ages, seasons and altitudes (all P<0.05). The serum level of 25-(OH)D and 25-(OH)D deficiency rate insufficient rate were (24.65±6.45) ng/L, 6.03%(128/2 122 cases) and 16.59%(352/2 122 cases), respectively.There were significant differences in the serum level of 25-(OH)D, 25-(OH)D deficiency rate and 25-(OH)D insufficient rate in children with different ages and seasons (all P<0.05). The mean serum Vit E level, Vit E deficiency rate and Vit E insufficient rate were (7.81±1.74) mg/L, 2.78%(59/2 122 cases) and 29.59%(628/2 122 cases), respectively.There were significant differences in serum Vit E level, Vit E deficiency rate and Vit E insufficient rate in children with different ages and seasons (all P<0.05). The mean serum levels of Vit A and Vit D remained the lowest before the age of 1 year, and their deficiencies at this age were the most significant.The mean serum level of Vit E remained the lowest in >1-2 years old, and its deficiency and insufficient at this age were the most significant.Vit A, D and E levels were significantly affected by seasonal changes, which were significantly higher in the summer than in the spring, autumn and winter.In addition, Vit A and 25-(OH)D were significantly affected by the altitude, which were the lowest above 4 km altitude. Conclusions:The overall serum levels of Vit A, 25-(OH) D and E in children with 0-6 years old in Tibetan Plateau areas of Ganzi Prefecture are lower than those in plain areas.Vit A, 25-(OH) D and Vit E levels significantly differed in the age, season and altitude, which are related to the lack of local resources, insufficient maternal nutrition during pregnancy and insufficient intake after birth, as well as temperature and light caused by changes in local seasons and altitude.Therefore, it is necessary to make reasonable supplements during pregnancy to prevent vitamin deficiency.