1.Establishment and evaluation of rabbit model of fecal incontinence.
Zong-Hai HUANG ; Xiao-Jing FU ; Zhou LI
Journal of Southern Medical University 2009;29(6):1170-1172
OBJECTIVETo establish and evaluate a rabbit model of fecal incontinence.
METHODSTwelve normal adult male New Zealand rabbits were randomly divided into experimental group and control group. The nerve innervating the external anal sphincter, namely the fourth sacral nerve, was functionally located and selectively damaged with local injection of 50 g/L ropivacaine in the experimental group, and normal saline injection was administered in the control group. The changes in the resting anal pressure was examined before and after the surgery, and the electromyogram (EMG) of the external anal sphincter was recorded for comparison with the pathological changes of the fourth sacral nerve.
RESULTSCompared with the control group, the experimental group exhibited significantly decreased resting anal pressure after the surgery. The EMG of the experimental group showed abnormal nerve conduction velocity of the fourth sacral nerve, suggesting successful nerve block. Transmission electron microscope revealed irreversible pathological changes in the ultrastructure of the axons of the fourth sacral nerve.
CONCLUSIONThis method allows successful establishment of fecal incontinence in rabbits, which facilitates further in vivo study of artificial sphincters for treatment of anal incontinence.
Amides ; administration & dosage ; Anal Canal ; physiopathology ; Animals ; Disease Models, Animal ; Electromyography ; Fecal Incontinence ; Lumbosacral Plexus ; Male ; Nerve Block ; Rabbits ; Random Allocation
2.Targeted killing of colorectal tumor cells by lentiviral constructs containing CD/TK suicide genes and KDR promoter.
Hai-jin CHEN ; Zong-hai HUANG ; Ai-guo WU ; Jin-long YU ; Guo-qiang SU
Journal of Southern Medical University 2007;27(5):624-627
OBJECTIVETo investigate the selective killing of colorectal tumor cells by lentivirus-mediated double suicide gene under the regulation of KDR promoter.
METHODS293T packaging cells were transfected with the plasmid FGW-KDRP-CD/TK to obtain the infectious viruses. KDR-expressing LoVo cells and LS174T cells that did not produce KDR were transfected with the recombinant virus, and the transfection efficiency was evaluated by the fluorecence microscope. RT-PCR was employed to examine the expression of CDglyTK. After treatment of the cells with 5-FC and GCV, the killing effects on the two cell lines were evaluated.
RESULTSThe recombinant construct showed similar infection rate of the two cell lines. RT-PCR demonstrated that CDglyTK gene was expressed only in LoVo cells infected with FGW-KDRP-CD/TK but not in LS147T cells, and the sensitivity of the two cell lines to the prodrugs was significantly different (P<0.001). The killing effect of the double suicide gene was much stronger than that of single suicide gene administered (P<0.001).
CONCLUSIONThe double suicide gene driven by KDR promoter has specific killing effect on the KDR-expressing colorectal tumor cells.
Antimetabolites ; pharmacology ; Apoptosis ; drug effects ; Cell Line ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Cytosine Deaminase ; genetics ; metabolism ; Flow Cytometry ; Flucytosine ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; Promoter Regions, Genetic ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Thymidine Kinase ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor Receptor-2 ; genetics
4.Selective cytotoxic effect of lentivirus-mediated double suicide gene transfer on human gastric adneocarcinoma cells.
Heng KONG ; Zong-hai HUANG ; Hai-jin CHEN ; Qiang LI ; Lin-yu TAO ; Ke QI
Journal of Southern Medical University 2010;30(1):47-50
OBJECTIVETo study the selective cytotoxic effect of lentivirus-mediated double suicide gene (CD/TK) against human gastric carcinoma cells SGC-7901 in vitro.
METHODSSGC-7901 cells were infected with FGW-KDRP-CD/TK vector and the infection efficiency was observed under a fluorescence microscope. The morphological changes of the infected cells were observed by Giemsa staining. Flow cytometry (FCM) was employed for cell cycle analysis, and the expression of CD/TK was detected by RT-PCR. The infected cells were then treated with the prodrugs ganciclovir (GCV) and/or 5-fluorocytosine (5-FC) at different concentrations, and the cytotoxic effects were evaluated using MTT method.
RESULTSThe infection efficiency of the lentiviral vector in SGC-7901 cells increased with the titer of the virus, which produced no significant effect on the cancer cell morphology in vitro or on the percentages of G0-G1, G2-M and S phase cells (P>0.05). RT-PCR demonstrated the expression of CD/TK gene in SGC-7901 cells infected by FGW-KDRP-CD/TK. The infected cells were highly sensitive to the prodrugs with a dose-dependent cytotoxic effect within a specific concentration range of the drugs, whereas the non-infected cells were not sensitive to the prodrugs. Combined use of the two prodrugs produced an obviously stronger inhibitory effect than either of the them (P<0.05). When combined, GCV and 5-FC at the concentration of 0.1+40, 1+80, 10+160, and 100+320 mg/L demonstrated a synergetic effect with a CDI<1.
CONCLUSIONLentivirus-mediated CD/TK fusion gene system can selectively kill gastric cancer cells, and the two prodrugs show a synergistic cytotoxic effect.
Adenocarcinoma ; genetics ; pathology ; Cell Line, Tumor ; Cytosine Deaminase ; biosynthesis ; genetics ; Cytotoxins ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Stomach Neoplasms ; genetics ; pathology ; Thymidine Kinase ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism
5.Clinical application of 64-slice computed tomographic angiography-based virtual colonoscopy in the diagnosis of colonic tumors.
Jin-long YU ; Zong-hai HUANG ; Chun-hui CUI ; Shu-xin HUANG ; Zhao-wei ZOU
Journal of Southern Medical University 2011;31(9):1585-1588
OBJECTIVETo investigate the clinical value of 64-slice computed tomographic angiography (CTA)-based virtual colonoscopy in the diagnosis of colonic tumors.
METHODSPhilips/Brilliance 64 CT volumetric scanning was performed in 8 patients with colonic cancer and 2 with colonic polypi identified by postoperative pathological examination. Mimics software was used for surface rendering of the intestine with the Marching Cubes algorithm for 3-dimensional (3D) virtual endoscope (VE) reconstruction and CTA-based 3D reconstruction of the large intestine and the surrounding structures. The location, volume and appearance of the lesions displayed by the virtual techniques were compared with the pathological results.
RESULTSThe 3D reconstruction was successfully completed in all the 10 cases, and the imaging diagnoses showed a total match with the pathological diagnoses. No significant differences were found between virtual endoscopy and CT virtual endoscopy. Virtual colonoscopy combined with digital model reconstruction provided valuable information for accurate identification of the position of the lesions and the complex adjacent anatomical structures.
CONCLUSIONVirtual colonoscopy based on 64-slice CTA, when combined with 3D reconstruction technique, allows accurate display of the colonic lesions and potential metastasis, which can be crucial for clinical staging and surgical planning of colonic cancer.
Adult ; Angiography ; methods ; Colorectal Neoplasms ; diagnostic imaging ; therapy ; Female ; Humans ; Image Processing, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Tomography, Spiral Computed
6.In vitro killing effect of adenovirus-mediated fusion gene system driven by KDR promoter on gastric cancer cells.
Guang-jun ZHOU ; Zong-hai HUANG ; Jin-long YU ; Zhou LI ; Guo-qiang SU
Journal of Southern Medical University 2006;26(4):402-405
OBJECTIVETo evaluate the killing effect of adenovirus(Ad)-mediated double suicide gene driven by kinase domain-containing receptor (KDR) promoter on gastric cancer MGC-803 cells.
METHODSThe 293 packaging cells were transfected by the plasmids pAdEasy-KDR-CDglyTK to generate infectious viruses. The gastric cancer MGC-803 cells were infected by the Ad followed by treatment with 5-FC and/or ganciclovir at different concentrations. The cell-killing effects were evaluated and the bystander effects analyzed after coculture of the cells without AdKDR-CDglyTK infection with the infected cells at different ratios. The cell cycle distribution was detected by flow cytometry and the pathological changes of the cells were observed by electron microscopy.
RESULTSThe infection rate of the resultant recombinant Ad in the cells increased gradually with increment of the multiplicity of infection (MOI) of the Ads. The killing effect of CD/TK fusion gene on the MGC-803 cells was much stronger than that of either of the single suicide gene (P<0.001), and considerable bystander effect was observed. The Ad infection caused MGC-803 cell growth arrest at G(1) phase with onset of apoptotic and necrotic morphologies of the cells as seen under electron microscope.
CONCLUSIONThe CD/TK fusion gene system driven by the KDR promoter possesses effective killing effect on the KDR-expressing gastric cancer MGC-803 cells.
Adenoviridae ; genetics ; Cell Line, Tumor ; Cytosine Deaminase ; genetics ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; Humans ; Promoter Regions, Genetic ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; Stomach Neoplasms ; pathology
7.Reconstruction of a digital three-dimensional model of the rectum and the surrounding structures based on CT angiographic data.
Jin-long YU ; Zong-hai HUANG ; Dong FU
Journal of Southern Medical University 2008;28(8):1466-1468
OBJECTIVETo reconstruct a digital three-dimensional model of the rectum and the surrounding structures based on CT angiographic (CTA) data.
METHODSBased on air pressure enema and CTA, the chest T12 level to upper portion of the femur of a healthy volunteer was scanned with 64-slice spiral CT in the arterial phase and venous phase. The rectum and the surrounding structures were reconstructed with Mimics software based on the two-dimensional images of 856 consecutive layers obtained by Dicom 3.0 standard CT. The model was validated using finite element analysis software.
RESULTS AND CONCLUSIONThe established three-dimensional digital model allowed clear visualization of such structures of the lumbar vertebrae, pelvis, femur, abdominal aorta, internal iliac artery, external iliac artery, branches of the external iliac artery, skin, rectum, the colons, part of the small intestines, and the urinary bladder and prostate. The application of thin-layer CT and Dicom 3.0 standard renders better accuracy of the established digital model, which can provide a platform for surgical skill training and teaching of anatomy.
Adult ; Angiography ; methods ; Aorta, Abdominal ; diagnostic imaging ; Finite Element Analysis ; Humans ; Iliac Artery ; diagnostic imaging ; Image Processing, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; methods ; Male ; Models, Anatomic ; Rectum ; anatomy & histology ; diagnostic imaging ; Tomography, Spiral Computed ; methods
8.Biological activity of survivin antisense oligonucleotide labeled with quantum dots or green fluorescein: a comparative study.
Hang YAO ; Zong-hai HUANG ; Zhou LI ; Rong HE ; Feng GAO ; Da-xiang CUI
Journal of Southern Medical University 2007;27(5):663-666
OBJECTIVETo compare the durability of quantum dots with that of green fluorescein for labeling survivin antisense oligonucleotide (ASODN) and investigate the difference in growth and apoptosis of cells transfected with the labeled survivin ASODN.
METHODSSurvivin ASODN labeled with quantum dots or green fluorescein was transfected into MCF-7 cells via Lipolifectmain(TM2000). The proliferation of MCF-7 cells was assessed with MTT assay, survivin mRNA expression determined by RT-PCR and its protein expression measured by Western blot analysis. The apoptosis rate of the transfected cells was estimated by flow cytometry, and the fluorescence distribution in the cells observed under fluorescent inverted microscope.
RESULTSThe mRNA and protein expressions of survivin were significantly decreased in the MCF-7 cells after cell transfection with survivin ASODN labeled with quantum dots or green fluorescein, and no significant difference was noted between the two labeling methods (P>0.05). Nor did survivin ASODN transfection with different labeling methods produced significant difference in cell proliferation and apoptotic rate (P>0.05). For green fiuorescein labeling, the fluorescence disappeared 4 days after transfection, whereas the fluorescence sustained for 1 week for quantum dots labeling.
CONCLUSIONSurvivin ASODNs labeled with quantum dots and green fiuorescein do not significantly differ in survivin expression or the transfected cell proliferation and apoptosis rate, but quantum dot labeling can be more stable with longer maintcnance of the labeling.
Apoptosis ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Fluorescein ; chemistry ; Gene Expression ; Humans ; Inhibitor of Apoptosis Proteins ; Microscopy, Fluorescence ; Microtubule-Associated Proteins ; genetics ; metabolism ; Oligonucleotides, Antisense ; chemistry ; genetics ; Quantum Dots ; Reverse Transcriptase Polymerase Chain Reaction ; Staining and Labeling ; methods ; Transfection
9.Phase IIa clinical trail of hemocoagulase acutus for injection.
Jun-jie ZHOU ; Zong-hai HUANG ; Jin-long YU ; Zhou LI ; Guang-jun ZHOU
Journal of Southern Medical University 2007;27(5):644-646
OBJECTIVETo evaluate the efficacy and safety of hemocoagulase acutus for injection and determine its curative dose.
METHODSForty-five patients on abdominal surgeries were randomly allocated into 2 study groups and 1 control group. Thirty minutes before the operation, the patients in the study groups received intravenous hemocoagulase acutus at 1 U and 2 U, respectively, and control group had no treatment. The hemostatic time, hemorrhagic volume, and hemoagglutination were observed in all the groups.
RESULTSThe average hemorrhagic volume and hemorrhagic volume per square were significantly lower in the two study groups than in the control group (P<0.05), and the average hemorrhagic volume per square were significantly lower in study group 2 U than in the 1 U group (P<0.05). No significant differences were found in adverse effects between the 3 groups.
CONCLUSIONHemocoagulase acutus for injection has good hemostatic effect for controlling capillary hemorrhage at the abdominal incisions and can be safely used in the surgical patients.
Abdomen ; surgery ; Adolescent ; Adult ; Aged ; Agkistrodon ; metabolism ; Animals ; Batroxobin ; administration & dosage ; therapeutic use ; Blood Coagulation ; drug effects ; Blood Loss, Surgical ; prevention & control ; Hemostasis, Surgical ; methods ; Hemostatics ; administration & dosage ; therapeutic use ; Humans ; Injections, Intravenous ; Male ; Middle Aged ; Treatment Outcome ; Young Adult
10.A short infrapectineal buttress plate fixing posterior column for complex acetabular fractures through Ilioinguinal ap-proach
Hu WANG ; Kun ZHANG ; Xing WEI ; Pengfei WANG ; Yuxuan CONG ; Zong LI ; Yahui FU ; Jinlai LEI ; Binfei ZHANG ; Hai HUANG ; Yan ZHUANG
Chinese Journal of Orthopaedics 2017;37(1):17-23
Objective To evaluate the clinical outcomes of ilioinguinal approach with short infrapectineal buttress plate fixing posterior column for patients with complex acetabular fractures. Methods Data of 29 consecutive patients (male 22, fe?male 7, age range 25-72 years, average age 53 ± 6.3 years) with complex acetabular who had operated by single ilioinguinal ap?proach with infrapectineal buttress plate from September 2008 to August 2012 were retrospectively analyzed. According to Letour?nel?Judet classifications, there were 4 cases of anterior column and posterior hemi?transverse, 11 cases of associated both?column, 4 of T?shaped and 6 with seagull sign. The acetabular fractures model was printed preoperatively by 3D printing technique for sur?gical simulation, open reduction and internal fixation through single ilioinguinal approach with a short infrapectineal plate fixing posterior column, particularly an ox horn shape K?wire sleeve was developed for drilling and screw insertion using flexible screw?driver. Clinical, radiographic, and functional outcomes assessed by the modified Merle d’Aubigné score were collected. Results Two patients were lost to follow?up, including one patient who died at 15 days postoperative because of pulmonary embolism, and the other one who had moved abroad at 12 months postoperative. The remaining 27 (93%) had a mean follow?up of 40 months (range, 24-75 months). The operating time was 180±35.5 min;the time for the fracture union was 3.5±0.9 month;blood loss during the operating was 500±43.9 ml;no case had fracture re?displacement. At the latest follow?up, radiographic grades were excellent in 17, good in 8, poor in 4, including one poor patient who had a total hip arthroplasty (3%) at 35 months after the internal fixation. The average modified Merle d’Aubigné score was 16 (range, 10-18), and categorized as excellent in 12, good in 8, fair in 5, and poor in 2. Three patients had developed deep venous thrombosis of the lower extremities, and one of them died of pulmonary embo?lism 15 days after operation; two cases of deep venous thrombosis of the lower extremity were found 3-4 days after operation. Thrombosis disappeared 6 months after anticoagulant therapy by warfarin. One case who had superficial wound infection after oper?ation was treated by anti?infection and VSD negative pressure, and incision was healed after one month. One case with poor quality of fracture reduction and seagull sign was treated with total hip arthroplasty after 35 months because of traumatic arthritis. The to?tal complication rate was 24%(7/29). Conclusion In the patient with complex acetabular fractures combining posterior column medial displacement, single Ilioinguinal approach with infrapectineal buttress plates could achieve a stable concentric hip joint, and immediate postoperative ambulation can result in reasonable clinical, radiographic, and functional outcomes.