2.Construction and Expression of The Recombinant of Hepatocellular Carcinoma-Targeting Adenovirus Containing r-Caspase-3 Gene
Wei WANG ; Wei CHENG ; Changru QIU
Chinese Journal of Bases and Clinics in General Surgery 2008;0(08):-
Objective To construct the recombinant of hepatocellular carcinoma-targeting adenovirus containing r-Caspase-3 gene and provide the gene therapic strategy for hepatocellular carcinoma. Methods The pAdTrack-EAFP-PALB was constructed and the r-Caspase-3 gene was subcloned into the vector. The linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183 cells. The candidate clone was analyzed by restriction endonuclease digestion and sequencing, and then pAdEasy-EAFP-PALB/r-Caspase-3 vector was digested with PacⅠand transfected into AD293 cells for packaging and amplifying, recombinant virus was constructed successfully. Infection titer and efficiency of recombinant virus were monitored by green fluorescent protein (GFP) expression. The expression of r-Caspase-3 in infected HepG2 cells was detected by RT-PCR and Western blot. The apoptosis of HepG2 cells was detected by SRB dyeing method. Results Shuttle vector pAdTrack-EAFP-PALB/r-Caspase-3 was correct after identification by restriction endonuclease analysis and sequencing. By PCR and PacⅠ restriction endonuclease analysis, the homologous recombinant of pAdEasy-EAFP-PALB/r-Caspase-3 was successful. The expression of GFP was observed when linearized pAdEasy-EAFP-PALB/r-Caspase-3 was transfected into AD293 cells. AD293 cells could be infected repeatedly by recombinant adenovirus. The expression of r-Caspase-3 gene on HepG2 cells was detected by RT-PCR and Western-blot methods respectively, which confirmed that the Ad-EAFP-PALB/r-Caspase-3 was constructed successfully. The specificity of Ad-EAFP-PALB/r-caspase-3 which targeting induced hepatocellular carcinoma cells was founded by SRB dyeing test. Conclusion The Recombinant of hepatocellular carcinoma-targeting adenovirus containing r-Caspase-3 gene was constructed successfully and which established the foundation of r-Caspase-3 gene therapy in future research to hepatocellular carcinoma.
3.Expression and significance of Ki-67 antigen in bladder cancer
Wei CHENG ; Jingyu WANG ; Dongwen WANG
Cancer Research and Clinic 2009;21(11):748-749
Objective To investigate the expression of Ki-67 in bladder transitional cell carcinoma and its relationship with grade and stage. Methods Immunohistochemistry method were used to detect the expression of Ki-67 in bladder transitional cell carcinoma and control groups. Results The positive expression rate of Ki-67 antigen in 60 cases of bladder cancer, 20 cases of benign bladder disease and 10 cases normal bladder mucosa were 25.9 %, 10.3 % and 1.1 %. There were significantly difference among each group. The more grade and stage, the greater expression of Ki-67 antigen. Conclusion Ki-67 antigen is related to cell proliferation. The expression of the Ki-67 antigen in bladder cancer is closely related to the grade extent of tumor, and it is an important cell proliferation indicator. As a signal, Ki-67 antigen reflects cell proliferation, measurement of the expression of the Ki-67 antigen could reflect the condition of tumor cell proliferation. It might be an important prognostic factor for judging the occurrence, development and prognosis of the tumor.
4.Selected three-field lymphadenectomy in thoracic middle-lower section esophageal carcinoma
Fuzeng WANG ; Cunshuan CHENG ; Yunfeng CHENG ; Guangqing WEI ; Qingliang WANG ; Zhibin CHENG ; Xiaogang CHENG ; Haiyun GUO
Journal of Chinese Physician 2011;13(1):53-57
Objective To explore the technique and effect of selected three-field lymphadenectomy by left thoracotomy in treatment of thoracic middle or lower section esophageal squamous carcinoma. Methods From Jun. 2005 to Mar. 2009, 213 patients with thoracic middle or lower section of esophageal carcinoma received esophagectomy, bilateral mediastinal lymphadenectomy and pleural membrane resection.Group 1 -5, 7 - 12a, 16al, and 19 were performed to dissect abdominal lymph node and extended thoracic and abdominal lymphadenectomy and only lymph node extraction of mesoesophagus in neck field. Results 14197 lymphatic nodes(LN) were detected in 213 case. The average number of resected LN was 66. 65 ±24. 73. The metastatic lymph node was detected in 105 cases. The metastatic rate was 49.05% (105/213).There were 423 metastatic lymph nodes. The lymph nodes metastasis was 2. 97% (423/14197) of all dissected lymphatic nodes. No remnant carcinoma in the upper and lower cutting edge was found in pathological examination. The operation time ranged from 2. 92 ~ 4. 67 ( 3. 37 ± 0. 42) hours. Blood transfusion during perioperative period was 0 ~ 6u ( 1.08 ± 0. 93 ) u. Perioperative plasma transfusion was 0 ~ 1400( 103.77 ± 184. 89) ml. The hospital-time was 14 ~ 39 ( 17.64 ±4. 12) days. There were no anastomotic leakage and recurrent laryngeal nerve injury. One case died from respiratory failure, the mortality was 0. 04% ( 1/213). Conclusion Surgical approach in the management of left thoracotomy in the sixth intercostals could extend resection of chest-field lymph node dissection, decrease neck field lymph node dissection. Abdomen-field lymph node dissection reached selected D3. The selected lymphadenectomy procedure had the advantages of small traumas and few complications.
5.Research progress of esophageal cancer mouse models
Wei MA ; Kai WANG ; Yufeng CHENG
Journal of International Oncology 2013;40(12):915-918
The esophageal cancer mouse models have three categories:chemical carcinogenesis model,xenograft tumor model and genetic engineering model.Chemical carcinogenesis model can simulate well the pathological processes of human esophageal cancer and is prepared simply.Xenograft tumor model is currently the most commonly used esophageal cancer animal model,which has advantages of rapid tumor formation,higher tumor formation rate and lower cost.Genetic engineering model involves mainly p53,Cyclin D1,Brca1 and p120ctn,and it helps to understand the biological characteristics of tumors,but it is expensive and the preparation time is long.
6.Differential gene expression and radiosensitivity of esophageal cancer
Kai WANG ; Wei MA ; Yufeng CHENG
Journal of International Oncology 2011;38(8):608-611
Radiotherapy is one of the most important treatment method for esophageal cancer. Cene expression profile of cancer cells may be one of key factors in determining the effectiveness of radiation therapy To provide guidance for individual therapy, research advances in gene expression involved in radiosensitivity of esophageal cancer are discussed from three aspects: radiation damage repair, cell cycle distribution and signal traneduetion.
8.Description and Comparison of Diagnostic Criteria in Childhood Functional Constipation
shu-cheng, ZHANG ; wei-lin, WANG
Journal of Applied Clinical Pediatrics 2004;0(07):-
Functional constipation(FC) in childhood is characterized by a low defecation frequency in combination with either involuntary loss of stools(encopresis),passage of large amounts of stool,retentive posturing,or hard and often painful defecation.It represents 3%-5% of general pediatric outpatient visits and up to 25% of pediatric gastroenterology consultations.FC is a diagnosis made by history and physical examination.No testing is necessary or desirable.The decision to seek medical care for symptoms arises from a parent′s or caretaker′s concern for children.The caretaker′s threshold for concern varies with his or her experiences and expectations,coping style,and perception of illness,till now there is not an universal native criteria established in spite of much has been done in it.The criteria used are still to draw assistance from the foreign standard,among which the classic criteria,the Rome Ⅱ and Rome Ⅲ criteria are considered commonly used.The purpose of this paper is to descript the several foreign criteria,and to make comparison among them,and to explore their applicability and effectiveness in childhood FC
10.Verapamil attenuates the progression of pacing-induced ion channel remodeling in atrial myocytes
Wei CHENG ; Yingbin XIAO ; Xuefeng WANG
Journal of Third Military Medical University 1988;0(05):-
Objective To study the potential effect of verapamil for the early changes of L-type calcium channel ?1c and potassium channel Kv4.3 in a rapid paced primary cultured atrial myocyte model for atrial fibrillation. Methods Primary rat atrial myocytes were cultured and established a rapid paced primary cultured atrial myocyte model. Atrial cells were divided into four groups: control group, rapid pacing group, verapamil with rapid pacing group and verapamil without rapid pacing group. Durations of rapid pacing were 24 h. The mRNA and protein expressions of L-type calcium channel ?1c and potassium channel Kv4.3 were measured by RT-PCR and Western blotting respectively. Results It was found that 24 h pacing significantly reduced mRNA and protein expressions of L-type calcium channel ?1c and potassium channel Kv4.3 as compared to controls (P0.05). Conclusion Expressions of L-type calcium channel ?1c and potassium channel Kv4.3 were reduced in the rapid pacing atrial myocytes, suggesting ion channel remodeling of atrial myocytes. However, verapamil can attenuate the progression of ion channel remodeling of atrial myocytes at least in early phase.