2.hG-CSF cDNA Cloning and the Construction and Applicaltion in Gene Therapy of Its Retroviral Vector
Weiping ZHANG ; Xuetao CAO ; Shihua MA ; Xin HUANG ; Minghui ZHNAG ; Qun TAO ; Tianxing YE ;
Chinese Journal of Cancer Biotherapy 1994;0(01):-
The hG -CSF cDNA was cloned by RT-PCR and confirmed by sequencing, which contains the full length of hG-CSF encoding region and parts of 5 '、3' non - coding region. Then the hG - CSF retroviral vector pLGSN was constructed by orientationally inserting the hG-CSF cDNA into the EcoRI/XhoI cloning sites of pLXSN vector. Packaged with CRE and CRIP packaging cell lines which are considered to be unlikely to produce helper viruses, the final pLGSN retrovirion titer reached 1. 1 ?106 CFU/ml. During constitutive passaging, the CRIP - LGSN cell clone produced relatively stable tilers of pLGSN retrovirion, ranging from 6. 8?105CFU/ml to 1. 1?106CFU/ml. By infecting the murine fibroblast cell line NIH3T3 with pLGSN retrovirion, a cell clone designated as NIH3T3 -G -CSF was obstained, secreting 168U/ml G-CSF . The integration and expression of hG-CSF gene in this cell clone were confirmed by Southern and Northern blotting analyses. Western blotting has also detected specifically the hG-CSF protein in the condensed supernalants from NIH3T3-G-CSF cells . After packaging the hG-CSF-secreting fibroblasts with collagen and implanting them into synergenic mice peritoneally , we detected a certain levels of G-CSF in the sera of mice, which suggested the implanted NIH3T3-G-CSF fibroblast cells could constitutively express and release hG-CSF in vivo. Our data showed the constructed hG-CSF retroviral vector could be used to further investigate the fibroblasl-mediated hG-CSF gene therapy .
3.Detection of coronary artery anomaly in patients with complex congenital heart disease by dual-source CT
Zhaoping CHENG ; Shihua ZHAO ; Huaibing CHENG ; Minjie LU ; Bin ZHAO ; Tao WANG ; Yanhua DUAN ; Lebin WU ; Ximing WANG
Chinese Journal of Radiology 2015;(7):515-519
Objective To explore the clinical usefulness of low-dose dual-source CT (DSCT) angiography in detecting coronary artery anomaly (CAA) in patients with complex congenital heart disease (CHD). Methods A total of 615 consecutive patients with complex CHD who underwent DSCT angiography between 2008 and 2012 were retrospectively reviewed. According to the ultrasonic results, a total of 312 patients with tetralogy of Fallot (n=176 cases) or double outlet right ventricle (n=72 cases) or pulmonary artery atresia (n=64 cases) were included. Scans were divided into 3 groups: retrospective electrocardiogram (ECG)-triggered spiral scanning in 75 cases, prospective ECG?triggered spiral scanning in 158 cases, and prospective ECG?triggered high?pitch spiral scanning in 79 cases. The subjective image quality was evaluated with a four?point scale by two radiologists. Radiation dose values were calculated. Interobserver agreement in subjective image quality grading was assessed by using Kappa statistics. The scanning groups were compared with the Kruskal?Wallis test. Results The incidence of CAA was 8.0 %(25/312). CAA was detected in 7.9%(14/176)patients with tetralogy of Fallot, 8.3%(6/72)patients with double outlet right ventricle, 7.8%(5/64)patients with pulmonary artery atresia. In 76%(19/25)of the patients, the anomalous vessels were crossing the right ventricular outflow tract (RVOT). Thirty of 312 CT examinations were nondiagnostic (9.6%). Two radiologists acquired good agreement (Kappa=0.72,P<0.01). The median subjective image quality score of three scan groups was 3 (range 1—4). The score in high?pitch spiral scanning group was significantly lower than those in other two groups (H=29.1,P<0.01). Retrospective ECG?triggered scans had a median dose of 1.19 mSv(range 0.52—3.29 mSv), prospectively ECG?triggered scans had a median dose of 0.51 mSv(range 0.27—2.13 mSv), and prospectively ECG?triggered high?pitch spiral scans had a median effective dose of 0.30 mSv(range 0.18—0.62 mSv). The difference between groups was statistically significant (H=160.0, P<0.01). Conclusions DSCT angiography is a reliable diagnostic method for the assessment of CAA with complex CHD and low?dose prospectively ECG?triggered DSCT scan is the best choice.
4.Changes in the number and biological ability of endothelial progenitor cells from peripheral blood of patients with systemic lupus erythematosus (SLE)
Peimei ZHOU ; Yonghong LU ; Xia XIONG ; Xiaoyun CHENG ; Yanjun ZHU ; Shihua ZENG ; Tao CHEN ; Qingbiao WA ; Qi CAI ; Qianming CHEN ; Cunhuo JIANG
Chinese Journal of Dermatology 2010;43(10):698-701
Objective To analyze the changes in number and biological ability of endothelial progenitor cells (EPCs) from peripheral blood of SLE patients. Methods Mononuclear cells (MNCs) were isolated by Ficoll density gradient centrifugation from peripheral blood of 20 female SLE patients and 20 healthy female controls. EPCs were identified by double staining using antibodies to CD34 and CD133, or antibodies to CD133 and vascular endothelial growth factor receptor 2 (VEGFR2). Phycoerythrin (PE) conjugated antiCD34, fluorescein isothiocyanate (FITC) conjugated anti-CD133 and APC conjugated anti-VEGFR2 antibodies were used in a three color flow cytometric analysis to determine the percentage of EPCs in peripheral MNCs.The proliferation and migration ability of EPCs were measured by MTT assay and modified millicell chamber assay, respectively. The adhesion activity of EPCs was evaluated by counting the number of adherent cells.Results The percentage and proliferation rate of EPCs in peripheral MNCs from female SLE patients were significantly lower than those from the healthy controls(4.49% ± 1.66% vs 20.81% ± 4.14%, 23.11% ± 3.16%vs 35.65% ± 1.74%, both P < 0.01 ). The migration and adhesion ability of EPCs from SLE patients was impaired compared with those from the healthy controls (12.00 ± 2.12 vs 23.60 ± 3.0 cells/field, 22.43 ± 4.43vs 36.43 ± 3.69 cells/filed, both P < 0.01 ). Conclusion There is a decrease in the number and an impairment in biological ability of EPCs in SLE patients.
5.Effect of intrathecal injection of morphine on patients with severe rectum cancer pain
Kejun DONG ; Shihua SUI ; Tao ZHAO ; Zhitao ZHAO ; Zhiqiang FA
Chinese Journal of Neuromedicine 2016;15(8):835-838
Objective To investigate the effect of intrathecal injection of morphine on patients with severe rectum cancer pain.Methods According to random number table method,28 patients with severe rectum cancer pain,admitted to our hospital from April 2014 to April 2015,were divided into two groups:intrathecal injection of morphine group (group Ⅰ) and epidural injection of morphine group (group E,n=14).Patients in group Ⅰ were treated with intrathecal injection of morphine,and group E with epidural injection of morphine;initial dose of group Ⅰ was 1/300 of daily average dosage,and group E 1/30 of daily average dosage;the additional dose of two groups was 1/10 of initial dose,with a locked time of 30 min.Resting and moving visual analogue scale (VAS) scores,life scores of quality of life questionnaire (QLQ)-C30,and daily dose of morphine were observed on the day before analgesia (T0),first week after analgesia (T1),second week after analgesia (T2),and first month after analgesia (T3).Nausea and vomiting,skin itching,retention of urine,and headache were recorded at the four points.Results As compared with those at T0,the resting and moving VAS scores and daily dose of morphine were significantly lower,life scores of QLQ-C30 at the T1,T2 and T3 were significantly higher in both two groups (P<0.05).As compared with those in group Ⅰ,the resting and moving VAS scores and daily dose of morphine in group E were significantly higher,life scores of QLQ-C30 in group E were significantly lower at the T1,T2 and T3 (P<0.05).The incidence of complications in group Ⅰ (21.4%)was significantly lower than that in group E (71.4%).Conclusion The effect of analgesia with intrathecal injection of morphine is better than that of analgesia of epidural injection of morphine in the treatment of severe rectum cancer pain.
6.Analysis and prospect of microwave ablation for hepatocellular carcinoma
Zeyu WANG ; Shihua HAO ; Lide TAO
International Journal of Surgery 2022;49(12):853-859
The global incidence of liver cancer has remained elevated for a long time. As a high-risk country for hepatitis B, China has one of the highest rates of liver cancer in the world. Historically, surgical excision has been the recommended method of treatment for early-stage liver cancer. With the advancement of imaging technology and the growing use of physical treatment in clinics in recent years, microwave ablation has emerged as a new treatment option for patients with small hepatocellular carcinoma and those who have missed the chance for surgery. It is less traumatic, requires less time in the hospital, and is less expensive than the traditional surgery. However, due to several current indication limits and the inability to totally avoid postoperative sequelae, microwave ablation is not appropriate for all patients with liver cancer. This article examines the use, combined therapy, and postoperative consequences of microwave ablation in the treatment of hepatocellular carcinoma, as well as the potential future direction of development in the treatment of hepatocellular carcinoma.
7.Application of 3D printing in surgical treatment of hepatobiliary malignancies
Shihua HAO ; Zeyu WANG ; Lide TAO
International Journal of Surgery 2023;50(4):285-288,F4
At present, surgical treatment is the most effective method for the treatment of hepatobiliary malignant tumor. However, due to the complex anatomical structure of hepatobiliary region, accompanied by vascular variation, and with the continuous update of medical concepts, the requirements for surgery are more strict. Traditional imaging examination has reached a bottleneck in the support of surgical treatment, while 3D printing technology is compared with the former. It showed strong advantages in preoperative program planning and improving the effect of intraoperative precise resection. At the same time, it also shows great potential for medical assistance and disease treatment in the production of bioactive models, and 3D printing technology has obviously enhanced the understanding of surgery for young doctors, and medical staff can create a variety of highly practical 3D printing models under the existing conditions. In the future, it is expected to overcome the limitations of materials and technology and bring higher therapeutic benefits for the majority of patients.
8.Changes of electroencephalogram of patients with drug resistant epilepsy after vagus nerve stimulation
Shihua DANG ; Lingli JIANG ; Haifeng SHU ; Tao YANG ; Zhi ZHANG ; Zhichuang QU ; Sixun YU ; Xin CHEN ; Yuan MA ; Yufang ZHAO ; Hongmei WANG
Chinese Journal of Neuromedicine 2022;21(9):912-915
Objective:To investigate the characteristics and change trends of electroencephalogram (EEG) in patients with drug resistant epilepsy (DRE) after vagus nerve stimulation (VNS).Methods:Twenty-five patients with DRE, admitted to our hospital from July 2016 to May 2019, were chosen; all patients accepted VNS and followed up for 12 months. Long range video EEG (VEEG) monitoring was performed before VNS, and 3, 6 and 12 months after VNS, and the tracing time of each monitoring was longer than 12 h. The EEG characteristics of these patients before and different times after VNS were analyzed.Results:In the VEEG monitoring before VNS, 25 patients showed sharp wave, spike wave, sharp slow wave, and compound spike slow wave in the interictal period; 3 patients (12%) could locate the brain region. The interictal EEG of 11 patients 3 months after VNS showed different degrees of improvement as compared with the preoperative one, which manifested as mixed rhythms: mono-spiking as sharp wave, sharp slow wave or spike wave; 8 patients had McHugh grading I-II. The interictal EEG of 18 patients 6 months after VNS showed different degrees of improvement as compared with the preoperative one; 11 patients had McHugh grading I-II. The interictal EEG of 21 patients 12 months after VNS showed different degrees of improvement as compared with the preoperative one; 15 patients had McHugh grading I-II.Conclusion:The EEG improvement effect of DRE patients after VNS is gradually improved with time; in some patients, the EEG improvement is earlier than improvement of clinical symptoms.
9.Changes of electroencephalogram of patients with drug resistant epilepsy after vagus nerve stimulation
Shihua DANG ; Lingli JIANG ; Haifeng SHU ; Tao YANG ; Zhi ZHANG ; Zhichuang QU ; Sixun YU ; Xin CHEN ; Yuan MA ; Yufang ZHAO ; Hongmei WANG
Chinese Journal of Neuromedicine 2022;21(9):912-915
Objective:To investigate the characteristics and change trends of electroencephalogram (EEG) in patients with drug resistant epilepsy (DRE) after vagus nerve stimulation (VNS).Methods:Twenty-five patients with DRE, admitted to our hospital from July 2016 to May 2019, were chosen; all patients accepted VNS and followed up for 12 months. Long range video EEG (VEEG) monitoring was performed before VNS, and 3, 6 and 12 months after VNS, and the tracing time of each monitoring was longer than 12 h. The EEG characteristics of these patients before and different times after VNS were analyzed.Results:In the VEEG monitoring before VNS, 25 patients showed sharp wave, spike wave, sharp slow wave, and compound spike slow wave in the interictal period; 3 patients (12%) could locate the brain region. The interictal EEG of 11 patients 3 months after VNS showed different degrees of improvement as compared with the preoperative one, which manifested as mixed rhythms: mono-spiking as sharp wave, sharp slow wave or spike wave; 8 patients had McHugh grading I-II. The interictal EEG of 18 patients 6 months after VNS showed different degrees of improvement as compared with the preoperative one; 11 patients had McHugh grading I-II. The interictal EEG of 21 patients 12 months after VNS showed different degrees of improvement as compared with the preoperative one; 15 patients had McHugh grading I-II.Conclusion:The EEG improvement effect of DRE patients after VNS is gradually improved with time; in some patients, the EEG improvement is earlier than improvement of clinical symptoms.
10.Application of boron-containing polyethylene as maze wall lining shielding for a 10 MV medical accelerator room
Yajing SONG ; Jing SU ; Shihua TAO ; Zechen FENG
Chinese Journal of Radiological Health 2025;34(1):73-76
Objective To evaluate the ambient dose equivalent rates of photons and neutrons inside and outside the door of a 10 MV accelerator room, and to report the shielding effect of boron-containing polyethylene as maze wall lining. Methods The ambient dose equivalent rates of photons and neutrons inside and outside the door of an accelerator room were taken as the research subject. The Kersey, Falcão, and modified Kersey methods were used to calculate the ambient dose equivalent rates of neutrons and neutron capture gamma rays inside and outside the door of the room before and after renovation. Measurements were made using an X-ray/γ-ray dose rate instrument and a neutron ambient dose equivalent rate meter. Calculated and measured results were compared. Results Before renovation, the measured neutron dose rate inside the door was 409 μSv/h, while the calculated values were 323 μSv/h (Kersey method), 428 μSv/h (Falcão method), and 219 μSv/h (modified Kersey method). The Falcão method yielded a value closest to the measured value, while the Kersey and the modified Kersey methods underestimated the value by 21% and 46%, respectively. After the installation of boron-containing polyethylene plates, the measured neutron dose rate inside the door was 190 μSv/h, with a 54% reduction. The neutron and photon ambient dose equivalent rates outside the door were 5.8 μSv/h and 6.0 μSv/h, respectively, before renovation, and 0.14 μSv/h and 1.6 μSv/h, respectively, after renovation. Conclusion For a 10 MV accelerator room, neutron shielding and protection measurements are necessary, especially for rooms with short mazes. The Falcão method provides the best estimate of neutron dose rates inside and outside the door. Using boron-containing polyethylene plates as maze wall lining is an economical and effective shielding method.