1.Construction of pDsVEGF165Red1-N1 and pIRES2-BMP2-EGFP and their co-expression using RFP and EGFP as reporter gene in HEK 293-T cells
Haibo ZOU ; Hong AN ; Dianming JIANG
Chinese Journal of Tissue Engineering Research 2006;10(21):174-176,封三
BACKGROUND: It has been proved that vascular endothelial growth factor 165(VEGF165) and bone morphogenetic protein 2(BMP2) can accelerate the vascularization synergistically.OBJECTIVE:To construct the vectors, pDsVEGF165Red1-N1 and pIRES2-BMP2-enhanced green fluorescent protein(EGFP) followed by co-transfected into HEK 293-T cells,and study their expression and location of VEGF165and BMP2 in the cells.DESIGN: A randomized and controlled experiment.SETTING: National Key Laboratory,Institute of Surgery Research,Daping Hospital,Third Military Medical University.MATERIALS: The experiment was conducted at National Key Laboratory,Institute of Surgery Research,Daping Hospital,Third Military Medical University from September 2002 to March 2004. pcDNA3.1\BMP2 ( gift of Dr.Bostrom, UCLA School of Mediine, Los Angeles,USA).pDsRed1-N1(gift of Pro. Roger Y.Tisen,University of California,San Diego,USA). pUC18/VEGF165,293-T cells(preserved by our Laboratory).METHODS: According to the nucleotide sequence of hVEGF165, the primers were designed.The hVEGF165 gene without stop codon was amplified by polymerase chain reaction (PCR).The fragment digested was cloned into the expression vector pDsRed1-N1.Meantime,the pIRES2-BMP2-EGFP expression vector was constructed.The two plasmids were co-transfected into HEK 293-T cells.The co-expression and distribution of the VEGF165and BMP2 were observed with confocal laser microscopy (CLSM) and detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blotting.MAIN OUTCOME MEASURES: Identification of the recombinant plasmids and the expressing mRNA and protein in 293-T cells.RESULTS: The recombinant plasmids were verified correct construction by restriction enzyme analysis, PCR and sequencing. The two genes which were co-transfected could express in HEK 293-T cells.The co-expression of the report genes,RFP and EGFP, were found over the cytoplasm and in the nuclei by CLSM.CONCLUSION: Two report gene expression vectors contain VEGF165 and BMP2 have been constructed successfully, which can be co-expressed in HEK 293-T cells. Thus, they can provide important and convenient tool to study intracellular interaction of VEGF165 and BMP2.
2.A study of single-walled carbon nanotubes modified by organics of the phthalocyanine category
Zhonghui XU ; Haibo NIU ; Yun JIANG
Journal of Pharmaceutical Analysis 2008;20(3):193-197
Organics of the phthalocyanine category have very good nonlinear optical properties. The single-walled carbon nanotubes were modified by using the phenoxy phthalocyanine. Characterization analysis was made by means of the transmission electron microscope (TEM), ultraviolet visible absorptive spectra, fluorescent spectra and Raman spectra. Under the TEM, it was observed that the composite looked like sugarcoated haws. By comparing the ultraviolet visible absorptive spectra before and after absorption, it was disclosed that the spectral intensity and the intensity of the peaks in the fluorescent spectra dropped remarkably. This shows that the single-walled carbon nanotubes have absorbed a large number of phenoxy phthalocyanines. Raman analysis revealed that in the Raman spectra, the position of the main peaks of the single-walled carbon nanotubes after absorption moved in the direction of long waves. The analysis suggests that the movement of the Raman spectra results from the change in the state of the single-walled carbon nanotubes before and after absorption.
3.Selection of internal fixation methods for treatment of upper segment complicated femoral fractures(USCFF)
Haibo ZHANG ; Qi JIANG ; Jia ZHENG
Orthopedic Journal of China 2006;0(04):-
[Objective]To investigate the best internal fixation methods of upper segment complicated femoral fractures(USCFF).[Method]Fifty-two cases(54 limbs) with USCFF were treated.In all patients open fracture reduction with use of internal fixation were done.Intramedullary interlocking nails(IIN) were used to treat 32 cases(34 limbs) of adult's USCFF and 130? plate were used to treat 20 cases of children's USCFF.Thirty-six limbs of them were closed fractures,and 18 limbs were opene.Measures of auto-ilium transplant(5 limbs),homolegous allograft bone transplant(10 limbs)were also taken.[Result]Fifty-two patients were followed-up from 9 to 40 months with an average of 16 months.Infection,fracture nonunions,malunion and femoral head necrosis complications were not found.The average period of union of adult and children fractures was respectively 6.8 months and 6.5 months.The long term effect was evaluated according to Ma Yuanzhang's evaluation standard,94.4 percent showed excellent function of joints and limbs.[Conclusion]Appropriate selection of internal fixation according to the age of patients and satisfactory fracture reduction are key points to improve outcome of USCFF.IIN in treating fiacture of adult and 130? plate in treating fracture of children are more ideal selections.
4.Long-term follow-up results of percutaneous balloon mitral valvuloplasty in mitral stenosis with severe pulmonary hypertension
Haibo HU ; Shiliang JIANG ; Ruping DAI
Journal of Interventional Radiology 1994;0(04):-
Objective To assess long term results (more than 5 year) after percutaneous balloon mitral valruloplasty (PBMV) on mitral stenosis (MS) with severe pulmonary hypertension. Methods Thirty patients after PBMV underwent critical evaluations including echocardiography, chest film and clinical status throughout the follow up period (6.4?1.4 years). Results Before and after PBMV and at follow up, mean mitral valve areas were (1.19?0.32) cm 2 vs (1.99?0.45) cm 2 vs (1.44?0.42)cm 2 respectively ( P
5.Value of CE-MRA Angiography in Diagnosis of Arteriosclerotic Occlusive Disease of Lower Extremity
Mingzhuang LIAO ; Haibo ZHAO ; Qitao JIANG
Chinese Medical Equipment Journal 2004;0(07):-
Objective To discuss the value of CE-MRA of lower extremity angiography in diagnosing Arteriosclerotic occlusive disease. Methods CE-MRA was performed on 25 patients with symptom of arteriosclerotic occlusive disease. CT angiograms were produced using maximal intensity projection. Results 24 examinations of CE-MRA were successful with 2 cases of straitness in right ailiaca communist, 2cases of straitness and 2 cases of occlusion in aorta abdominal, 4 cases of straitness and 2 cases occlusion in artery of femoral, 3 cases of straitness and 4 cases of occlusion in artery of crus. One case was failed for the patient himself. Conclusion CE-MRA of lower extremity angiography can monitor multiple arteries of Arteriosclerosis occlusive disease of lower limbs and observe the extent and range of lesions. It has high diagnostic accuracy in patients with lower extremity artery diseases, and it is a safe, no-trauma and effective method in lower extremity artery.
6.Comparison of two different kinds of primary hydroxyapatite orbital implant placement
Qian XIANA ; Xueliang XU ; Haibo JIANG
Chinese Journal of Tissue Engineering Research 2008;12(26):5185-5188
BACKGROUND: There are two commonly used techniques of primary orbital implant placement, one is using sclera to cover hydroxyapatite prosthesis after evisceration and the other is using sclera to wrap around the hydroxyapatite prosthesisafter enucleation, which one is more efficient and safer.OB.IECTIVE: To compare the clinical safety of the two kinds of surgery for placing hydroxyapatite prosthesis in the orbit.DESIGN, TIME AND SETTING: A retrospective case analysis was performed.between June 2000 and June 2005 at Xiangya Hospital, Central South University, Changsha, Hunan Province, ChinaPARTICIPANTS: Fifty eyes of 50 cases who were admitted between June 2000 and March 2003 to accept enucleation implants, including 39 cases with atrophic eyeball, 6 cases with anterior scleral staphyloma, 1 case with absolute glaucoma, 4cases with eyeball rupture. Forty-nine eyes of 49 cases who were admitted between April 2003 and June 2005 to accept evisceration implants, including 36 cases with atrophic eyeball, 8 cases with anterior scleral staphyloma, I case with absolute glaucoma, 4 cases with eyeball rupture. All the operations performed by the same surgeon.METHODS: In enucleation implant group, autologous sclera-wrapped hydroxyapatite orbital implant was placed into the muscle cone after enucleation. In evisceration implant group, the bared hydroxyapatite orbital implant was placed into muscle cone after evisceration, and was covered by autologous sclera in anterior.MAIN OUTCOME MEASURES: Appearance improvements and complications of patients after orbital implant placement. RESULTS: There was no significant difference between the two groups in appearance improvement. In enucleation implant group, 7 patients experienced conjunctival dehiscence and 5 patients experienced orbital implant exposure. In evisceration implant group, no conjunctival dehiscence and orbital implant exposure occurred. There were significant differences between the two groups in conjunctival dehiscence ( x2 =7.380, P < 0.01) and orbital implant exposure( x2 =5.160, P < 0.01 ).CONCLUSION: Evisceration with sclera-covered orbital implant is more simple, safe, convenient and effective than enucleation with sclera-wrapped orbital implant.
7.The antiangiogenic effect of thalidomide on murine liver metastases
Hong LI ; Xuyuan JIANG ; Haibo SHAO
Chinese Journal of Radiology 2009;43(6):656-660
Objective To investigate the morphological change of intratumoral microvessels after administration of thalidomide in the murine hepatic metastases. Methods Among 20 mice with hepatic metastases created by injection of colon-26 tumor cells into the spleen, 10 were treated with thalidomide (200 mg/kg) by intraperitoneal injection daily, the other 10 were treated with saline only by intraperitoneal injection daily. Fifteen days after inoculation of tumor cell, the intratumoral mierovessel of hepatic metastases with similar size in both groups were studied with in vivo microscopy (26 and 27 neoplasms in experimental group and control group respectively ) and immunohistochemistry for CD34 (52 and 55 neoplasms in experimental group and control group respectively). Two-tailed student t test was used to determine differences in intratumoral microvessel density (MVD), intratumoral branch density (BD) and CD34 positive intratumoral microvessel density (MVD-CD34) between the small ( < 400 μm in diameter) and large metastases in both groups, and that between thalidomide treated group and control group. Results For the control group, although the MVD and MVD-CD34 of larger metastases was more than that of small metastases respectively [(18.1±3.5)% vs (13.0±3.2) %, t =2.840,P<0.01;(22.9±2.8)vs ( 12. 8±2. 5) vessels per field, t = 9. 860, P < 0. 01 ], the BD was similar to that of small metastases [(110.0±20.5)vs(99.7±17.3) branches/rnm2, t = 1.040,P >0.05]. For the thalidomide treated group, despite the MVD-CD34 of larger metastases was more than that of small metastases [ ( 17.4±2. 3)vs (11.5±2. 5 ) vessels per field, t = 8. 770, P < 0. 01], the MVD and BD was similar to that of small metastases respectively [(14.7±3.5)% vs(13.2±3.3) %, t =0.826,P >0.05; (95.3±18.3)vs (97. 1±21. 0)branches/mm2,t=0. 347,P>0. 05]. The MVD, BD and MVD-CD34 of small metastases were similar to each other between two groups ( t = 0. 098, 0. 392,1. 190; P > 0. 05 ), however, that of large metastases were significantly lower in thalidomide treated group than in control group ( t = 3. 140,2. 870, 9. 850;P < 0. 01 ). Conclusions Thalidomide exerts antiangiogenic effect on the hepatic metastases with angiogenesis only, and the different vascular components in the tumor vasculature demonstrate variousresponses to antiangiogenic therapy.
8.Application of moist electrothermal coagulation for hemostasis in iatrogenic splenic injury
Haibo CHEN ; Yinghao JIANG ; Chengfei SHI
Chinese Journal of General Practitioners 2021;20(3):359-361
The iatrogenic splenic injuries occurred in 49 patients who underwent abdominal surgery from January 2005 to January 2020; including grade I injury in 40 cases and grade Ⅱ injury in 9 patients. The normal saline-soaked gauze was placed on the wound surface of the injured spleen, then the high-frequency electric coagulator was employed for hemostasis. The bleeding was successfully stopped and the spleens were preserved without postoperative complications in all cases. The results show that moist electrothermal coagulation can be effectively and safely applied in treatment of grade I and grade Ⅱ injury iatrogenic splenic injuries, and no special equipment required.
9.Application of severing splenomental fold priorly to prevent iatrogenic splenic injury in laparoscopic radical gastrectomy
Haibo CHEN ; Yinghao JIANG ; Chengfei SHI
Chinese Journal of General Practitioners 2021;20(5):598-601
A total of 521 patients underwent laparoscopic radical gastrectomy from January 2013 to January 2020 in the First People′s Hospital of Wenling. In 242 cases the splenomental fold was severed before dissecting the left half of omentum or spleen (pretreatment group), and in 279 cases the splenomental fold was not severed priorly (routine group). For pretreatment group the introoperative splenic injury occurred in 4 cases (1.65%), including 3 cases (1.24%) with class Ⅰ injury and 1 case (0.41%) with class Ⅱ injury; while for routine group splenic injury occurred in 24 cases (8.60%), including 22 cases (7.89%) with class Ⅰ injury and 2 cases (0.72%) with class Ⅱ injury, and the rupture of splenic capsule caused by tracting splenomental fold occurred in 19 cases(6.81%). There were significant differences in total number of splenic injuries, splenic injuries with class Ⅰ and rupture of splenic capsule caused by tracting splenomental fold between two groups ( P<0.05). The operation time of 28 cases with splenic injuries was (185±89) min, which was longer than that in 493 cases without splenic injuries [(172±95) min, P<0.05]. The results show that rupture of splenic capsule by tracting splenomental fold is main cause of splenic injury in laparoscopic radical gastrectomy and most of them are class Ⅰ injuries. To sever the splenomental fold priorly can reduce the incidence of iatrogenic splenic injury.
10.The comparative study of IOL Master and A ultrasound combined with corneal topography measurement for IOL power
Kehua WANG ; Jianzhou WANG ; Jian JIANG ; Haibo JIANG ; Xiaobo XIA
Journal of Chinese Physician 2014;(4):450-455
Objective To investigate the difference of IOL Master and A-ultrasound combined with corneal topography meas-urement in intraocular lens ( IOL) power calculation .Methods A total of 84 eyes of 60 cataract patients received phacoemulsification and intraocular lens implantation surgery were selected in Xiangya Hospital from March 2010 to September 2011.There were 57 eyes of 42 age-related cataract patients and 27 eyes of 18 complicated cataract patients .The patients were divided into IOL Master group and A ultrasound combined with corneal topography measurement group .Before surgery , in IOL Master group , axial length ( AL) and corneal curvature were measured with IOL Master , IOL power was calculated according to the Haigis , SRK/T formula and so on.At the same time the axial length ( AL) and corneal curvature were measured with A ultrasound combined with corneal topography respectively and IOL power was calculated by SRK/Ⅱ formula.In A ultrasound combined with corneal topography measurement group , axial length ( AL) and corneal curvature were measured with A ultrasound combined with corneal topography respectively , IOL power was calculat-ed according to the SRK/Ⅱformula.3 months postoperatively , all the patients were conducted refractive outcome and calculating mean absolute refractive error(MAE).Finally the data were analysed.Results ⑴In IOL Master group, before operation there was no sig-nificant difference in mean axial length between IOL Master and A-ultrasound measurement ( P >0.05 ) , while in the patients with AL>26 mm the axial length was (28.53 ±0.57)mm and (29.42 ±0.64)mm using IOL Master and A-ultrasound measurement respec-tively ( P <0.05).The mean corneal curvature was (42.12 ±0.31)D and (43.09 ±0.27)D using IOL Master and corneal topogra-phy measurement respectively ( P <0.01).The mean IOL power were (17.06 ±0.48)D and (16.37 ±0.56)D in IOL Master group and A ultrasound combined with corneal topography measurement group respectively ( P <0.05 ) .⑵3 months postoperatively , the MAE was (0.07 ±1.05)D and (0.16 ±0.81)D in IOL Master group and A-ultrasound combined with corneal topography measure-ment group( P <0.05) respectively.Conclusions In cataract patients with normal axial length , IOL Master and A-ultrasound com-bined with corneal topography have high consistency for axial length measurement and IOL power calculation .But in cataract patients with high myopia , IOL Master is more accurate for axial length measurement and IOL power calculation compared to A -ultrasound com-bined with corneal topography .