1.Effects of mifepristone on the growth of human endometrial carcinoma cell line HHUA xenografted in nude mice of and the expressions of COX-2,CDK4
Hongguo LI ; Xingbo ZHAO ; Yan MENG
China Oncology 1998;0(04):-
Background and purpose:Endometrial carcinoma is a common malignant tumour in women and expresses COX-2,CDK4;the expressions are associated with the development of the tumour. Mifepristone can inhibit the growth of the tumours, but its mechanisms are not very clear, We investigated the inhibitory effect of antiprogestins mifepristone n the growth of human endometrial carcinoma cell line HHUA xenografted in nude mice of and the expressions of COX-2,CDK4,the purpose was to find out whether or not mifepristone could influence the expressions of COX-2,CDK4. Methods:Human endometrial carcinoma HHUA cells were cultured in vitro.The models of xenografted tumor were established by the transplantation of human endometrial carcinoma HHUA cell on nude mice. The nude mice were randomly divided into two groups to receive either refined peanut oil (control group), or daily mifepristone for six weeks, respectively . The sizes of xenograft were measured in the pre-and post-treatment. The changes of morphology were observed by glass microscopy. The positive immunostaining for COX-2 and CDK4 were evaluated semiquantitatively by using an immunohistochemical scoring system (HSCORE) that it incorporated both the intensity and the distribution of specific staining.Results:After 6 weeks of treatment , the size of the xenografts in MIF group were 115.25?10.97 mm 3 ,compared with 313.25?43.92 mm 3 in the control group (P
2.A Practical and Efficient Method for The Retrieval of Ancient DNA Sequence
Ranran LIU ; Jing YUAN ; Xingbo ZHAO ; Ning LI
Progress in Biochemistry and Biophysics 2009;36(11):1495-1502
Retrieval of ancient DNA (aDNA) sequences from organism remains provide direct view of their evolutionary history. However, researches on aDNA have suffered from lots of technical problems. Specifically, discredited sequences were generated from damaged aDNA templates, and expensive and time-consuming methods were employed. Here, a method which could recover the endogenous aDNA as well as to reduce the cost and research period is described. This is achieved by improving the ancient DNA extraction method of isopropanol precipitation, and reevaluating the method of PCR after N-glycosylase (UNG) treatment, which could remove the damaged DNA from the aDNA extract. The efficiency of these methods were tested by comparing with traditional methods using ancient specimens of pig teeth aged between 4 300 years before present (BP) and 3 900 BP. The results showed that: firstly, the extraction efficiency of the improved method of isopropanol precipitation and current method with silica-based spin column were all 60%. Furthermore, the research period at least could be reduced by half with the application of the improved methods and the cost to 1/10 of the current method. Secondly, sequences obtained through the method of PCR after UNG treatment were 100% authentic. In contrast, 66%~ 88% sequences were authentic based on the results obtained with the method of multiple PCRs without UNG treatment. And the research cost and period needed by the method with UNG treatment were only half of the later one. These results demonstrate that the improved extraction method of isopropanol precipitation combined with the method of PCR after UNG treatment could increase the success rate of authentic DNA amplified and at least reduce the research cost and period by half. Therefore, this method can be applied in the large-scale detection of ancient specimens.
3.Study of effect of mifepristone on apoptosis of human ovarian cancer cell line 3AO
Qiang LI ; Jijun LI ; Xingbo ZHAO ; Mei JI
Chinese Journal of Obstetrics and Gynecology 2000;0(10):-
Objective To investigate the effect of mifepristone on the activity of proliferation and the apoptosis, the expression of estrogen receptor (ER),progesterone receptor (PR) protein and morphology changes of human ovarian carcinoma cell line 3AO and SKOV3 in vitro. Methods The proliferative activity of 3AO and SKOV3, which were cultured in vitro, was measured by tetrazolium-based colorimetric assay (MTT assay). Flow cytormetry (FCM) was used to measure the expressive rate of ER, PR , p53 protein, bcl-2 protein, cell apoptotic rate and cell proliferative cycle of 3AO cells, which were cultured with different concentration and duration of mifepristone. The morphologic and ultrastructure changes of apoptotic 3AO cells was observed by the light and electron microscopy. Results Mifepristone inhibited significantly the proliferation of 3AO cells in dose-time dependent manner in vitro. The inhibitory rate of 3AO cells growth, which were cultured with different concentration of mifepristone( 5,10,20,40,80 ?mol/L) and duration (24,48,72 h) was from 1.7% to 75.0%(P0.05). 3AO cells apoptosis activity appeared the positive correlation with the dose of mifepristone and cultured duration (P
4.Bcl-2 associated athanogene 3 affects the epithelial-mesenchymal transition in human cervical cancer
Li WEI ; Xiaopeng QIN ; Xingbo ZHAO ; Wei WANG
Chinese Journal of Obstetrics and Gynecology 2017;52(8):551-557
Objective To investigate the expression of Bcl-2 associated athanogene 3 (BAG3) in cervical cancer tissues and cells and its role in epithelial mesenchymal transition (EMT) of cervical cancer.Methods (1) Cervical cancer samples were collected from September 2015 to March 2017 in the Qilu Hospital of Shandong University and Shangdong Provincial Hospital.While,50 normal tissues were collected from August 2015 to March 2017 in the Dezhou Municiple Hospital,which were obtained from patients with uterine mnyoma underwent hysterectomy and patients with cervical biopsy.Reverse transcription (RT)-PCR and western blot were used to detect the expression of BAG3 mRNA and protein,and their clinical significances were analyzed.(2) The expression of BAG3 mRNA and protein was detected using RT-PCR and western blot method in HeLa and SiHa cell lines and normal cervical epithelial cells.The experiment was divided into two groups,BAG3 small interfering RNA transfected group (st-BAG3) and the control group transfected with small interfering RNA (siRNA).Cell counting kit 8 (CCK-8) analysis was used to detect cell proliferation of two groups.Wound-healing and transwell assay were used to detect the migration and invasion ability of HeLa and SiHa cells.The xenograft model of cervical cancer in nude mice was used to observe the effect of BAG3 on tumor xenografts and the tumor-related biomarkers were tested by western blot.Results (1) The expression levels of BAG3 mRNA and protein in cervical carcinoma tissues were 1.20±0.15 and 1.10±0.16,which were significantly higher than that in normal cervical tissue,0.23± 0.04 and 0.29 ± 0.03 (both P<0.01).The results showed that the expression levels of BAG3 mRNA and protein were significantly correlated with cervical carcinoma staging and lymph node metastasis (P<0.05).However,its expression was not conrelated with the patient's age,pathological grade,and diameter of tumor (all P>0.05).(2) Compared with normal cervical epithelial cells,the expression of BAG3 mRNA and protein levels in HeLa and SiHa cells were significantly increased (P<0.01),the expression levels of BAG3 mRNA and protein in HeLa and SiHa cells transfected with si-BAG3 were significantly lower than that in control group (all P<0.01).After post-transfected 72 hours,A value of HeLa and SiHa with transfection were significantly lower than those in control group [(0.88±0.08) vs (1.22±0.13),(0.92±0.09) vs (1.35±0.12);both P<0.01].After post-transfected 24 hours,the migration level of HeLa and SiHa cells with transfection were significantly lower than those in the control group [(20.1±2.1)% vs (58.6±5.6)%,and (21.1±2.1)% vs (61.7± 5.4)%;both P<0.01].The transmembrane cell number in HeLa and SiHa cells with transfection were 76± 11 and 71±8,which were significantly less than those in control group (131± 12 and 129± 14;both P<0.01).After the inoculation into nude mice,tumor formation time of HeLa and SiHa cells with transfection were (9.5±0.5) and (10.5 ± 1.3) days,respectively,which were significantly longer than those in control group [(4.5±0.5) and (5.2± 1.1) days;both P<0.05].Compared with those in the control group,the expression level of Slug,N-cadherin and matrix metalloproteinase-2 (MMP-2) protein in HeLa and SiHa cells with transfected in tumor tissues were significantly decreased (all P<0.01),while the expression level of E-cadberin protein was significantly increased (P<0.01).Conclusion BAG3 could be involved in the proliferation,migration and invasion of cervical cancer cells by affecting cervical cancer EMT,and BAG3 may be an effective target for the treatment of cervical cancer.
5.Inhibition effect of exemestane in endometrial carcinoma cell line HHUA in vitro
Hongguo LI ; Xingbo ZHAO ; Mingjiang LI ; Jijun LI
China Oncology 2001;0(02):-
0.05),while those of the cells treated with 6?10-5,6?10-6,6?10-7 mol/L of exemestane were significantly different from that of controls(P
6.The experimental of the optimal angle position of X-ray in the dosed reduction of femoral neck fracture using internal fixation
Fei LI ; Jianxiong MA ; Xingbo WANG ; Xuan JIANG ; Xingwen ZHAO ; Biao HAN ; Ying WANG ; Bin LU ; Xinlong MA
Chinese Journal of Orthopaedics 2017;37(12):735-745
Objective To find out the display rules of the key part of the proximal femur and fracture line and obtain the best viewing position and angle by placing the femoral neck at different positions and different angles through X-ray.Methods Six dry specimens of cadaveric femur were from Department of Anatomy,Tianjin Medical University.Three models of complete femoral neck model,tin line fracture model and steel saw fracture model were made respectively.The tin line fracture model was based on the Pauwells angle,using the tin wire(1mm) wrapped around the femoral necks to make three kinds of fracture models (Pauwells angle 30°,50° and 70°);steel saw fracture model was made by hacksaw,then reposition in situ,to make three kinds of fracture models same as the tin line fracture model.The projection manner included different positions and different angles,different projection positions include:parallel with the femoral shaft,perpendicular to the femoral shaft,parallel to the femoral neck and perpendicular to the femoral neck;different projection angles included:from 40°,30°,20°,15°,10° and 5° in the head side to 5°,10°,15 °,20 °,30 °and 40° in the foot side and vertical angle 0°.For the complete femoral neck model,we observed the imaging characteristics of the key parts of the femoral head and neck (tension trabecular bone and pressure trabecular bone;lesser trochanter;intertrochanteric line;length of neck of femur and femoral head shape),looking for the display rules,and obtained the best viewing position.For tin line fracture model and steel saw fracture model,we tried to find the best display angle and position by the different projection position and angle.Results The results of complete femoral neck model:Lesser trochanter:in perpendicular to the femoral shaft position showed the best,gradually increased with the foot side deflection;tension trabecular bone and pressure trabecular bone:in parallel with the femoral shaft position 10°on the head side;intertrochanteric line:no difference between parallel with the femoral shaft with perpendicular to the femoral shaft;Femoral neck shape:deflect to both sides,head of femur was out of shape.The results of tin line fracture model:the Pauwells angle 30° model showed the best position in parallel with the femoral shaft position 20° on the head side;the Pauwells angle 50° model showed the best position in parallel with the femoral shaft position 5° on the head side;the Pauwells angle 70° model showed the best position in parallel with the femoral shaft position 10° on the head side.The results of steel saw fracture model were the same with the tin line fracture model.Conclusion There was the best viewing angles and positions for the key anatomy of the proximal femur and different Pauwells angle classification of femoral neck fracture.The image doctor could make more accurate projection,according to the different types of femoral neck fracture.
7.Microscopic authentication method of traditional Chinese medicine Gusuibu.
Yingxia ZHU ; Bengang ZHANG ; Yaodong QI ; Haitao LIU ; Zhao ZHANG ; Xingbo LI ; Shanshan ZOU
China Journal of Chinese Materia Medica 2012;37(8):1152-1156
OBJECTIVETo provide practical method for microscopic authentication of traditional Chinese medicine Gusuibu and its adulterants.
METHODBy means of light microscope, scanning electron microscopy and tissue section techniques, the morphology, the size of the rhizome scales and their bearing position in the original plants of Gusuibu and its adulterants, i. e. Drynaria roosii, D. delavayi, D. quercifolia and Pseudodrynaria coronans were analyzed.
RESULTThere were significant differences between scales length of D. roosii, D. delavayi and P. coronans, while there was no significant difference between that of D. roosii and D. quercifolia. The scale teeth of D. delavayi were usually curved, bifid and uneven distributed at the scale fringe, which was different from that of the other three species. The base of the scales sinks in epidermis in D. roosii, D. quercifolia, and P. coronans, while it bore at the raised part of epidermis in D. delavayi.
CONCLUSION[corrected] Morphology, size and bearing position of the rhizome scales have significant differences in the several species. Therefore, these characteristics can be applied to the identification of Gusuibu and its adulterants.
Medicine, Chinese Traditional ; methods ; Microscopy ; Microscopy, Electron, Scanning ; Polypodiaceae ; anatomy & histology ; classification ; Rhizome ; anatomy & histology ; classification
8.Free thin sensate superficial circumflex iliac artery perforator flap based on lateral cutaneous branch of the subcostal nerve for repair of soft tissue defect in the foot and ankle
Zhaohui PAN ; Shan XUE ; Yuxiang ZHAO ; Hongfei LI ; Peng GAO ; Jianbo WANG ; Xingbo LI ; Jianli WANG
Chinese Journal of Trauma 2021;37(6):526-531
Objective:To investigate the efficacy of free thin sensate superficial circumflex iliac artery perforator flap based on lateral cutaneous branch of the subcostal nerve for repair of soft tissue defect in the foot and ankle.Methods:A retrospective case series study was made on clinical data of 9 patients with soft tissue defect in the foot and ankle admitted to 80th Group Military Hospital from December 2017 to December 2019. There were 8 males and 1 females, with the age of 28-63 years [(47.3±12.3)years] and the body mass index (BMI) of 16.7-27.8 kg/m 2 [(23.9±3.9)kg/m 2]. The size of soft tissue defect ranged from 10 cm×6 cm to 20 cm×12 cm. All patients were treated with the free thin sensate superficial circumflex iliac artery perforator flap based on lateral cutaneous branch of the subcostal nerve. After debridement or tumor resection, a thin sensate flap was harvested by intra-adipose tissue dissection between the superficial and deep fat layers. The size of flap ranged from 13 cm×8 cm and 13 cm×10 cm. The thickness of the defatted flap ranged from 3-6 mm. The distance from the anterosuperior iliac spine to the point where the lateral cutaneous branch of the subcostal nerve crossed the iliac crest ranged from 7.5-10.0 cm. The flap survival, complications, and reoperation were observed after operation. The sensory recovery of the flap was evaluated using Tinel sign and nine-grid method including monofilament touch perception, vibration perception, pinprick perception, temperature perception, and static two-point discrimination test. The joint range of motion, and shoewear and walking problems were recorded. At the last follow-up, the American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score was used to assess the affected foot and ankle. The injury at the donor site was detected as well. Results:All patients were followed up for 6-35 months [(21.1±10.1)months]. All flaps survived without infection or tumor reoccurrence. One patient developed ulceration, then surplus skin on the reconstructed heel was resected. One patient underwent flap debulking and removal of internal fixation. One or more sensory modalities within the nine areas in each flap could be detected at postoperative 3-6 months. The monofilament touch, vibration, pinprick, and temperature perception were presented in almost all regions of each flap at postoperative 12 months. However, only one patient in one region was noted with the static two-point discrimination, in which the distance of the two points was set as 25 mm. The range of ankle motion was slightly limited in 2 patients who underwent osseoligamentous complex reconstruction. All patients were able to wear normal shoes and walk without pain. At the last follow-up, the AOFAS ankle-hindfoot score ranged from 78 to 97 points [(86.4±7.4)points], significantly improved from preoperative 10-70 points [(44.2±18.4)points] ( P<0.01). No patients complained of pain at the donor site, but the widening linear scar was noted. Conclusion:For medium-sized soft tissue defect of the foot and ankle, the free thin sensate superficial circumflex iliac artery perforator flap based on lateral cutaneous branch of the subcostal nerve can be defatted with the requirement and has advantages in defect site appearance, sensory restoration, wearing ordinary shoes, painless walking, good functional recovery, and minimal donor site morbidity.
10.Flap combined with 3D printed microporous tianium(tantalum)prosthesis in the treatment of lower extremity soft tissue defect with large bone defect
Yongqing XU ; Xinyu FAN ; Teng WANG ; Shaoquan PU ; Chuan LI ; Xingbo CAI ; Yi CUI ; Xiaoqing HE ; Wei LIN ; Yipeng WU ; Jian SHI ; Xia LI ; Dewei ZHAO ; Baoyi LIU ; Qiang FENG
Chinese Journal of Microsurgery 2022;45(1):21-27
Objective:To investigate the effect of flap combined with 3D printed microporous titanium(tantalum)prosthesis in the treatment of lower extremity soft tissue defect with large bone defect.Methods:From January 2019 to December 2020, 2 patients with large soft tissue defects on dorsal foot together with large metatarsal bone defect and 4 patients with soft tissue defects of calf with large tibial bone defect were treated. The areas of soft tissue defect were 5.0 cm×8.0 cm-15.0 cm×10.0 cm. The length of the bone defect were 3.8 cm to 7.0 cm, 5.75 cm in average. In the first stage, metatarsal bone defect or tibial bone defect was filled with vancomycin blended bone cement, meanwhile, soft tissue defect was repaired with anterolateral femoral flap(ALTF) with vascular anastomosis in 2 cases of feet, and local fascia flap was trans-positioned in 4 cases of lower extremity defects. The sizes of repairing flap were 6.0 cm×8.5 cm-16.0 cm×11.0 cm. Two to 7 months after the initial surgery, the customer designed microporous titanium prostheses were used(5 cases with microporous titanium and 1 with microporous tantalum) to repair the bone defects. The wound healing, the integration of metatarsal and tibial fractures with 3D printed microporous titanium(tantalum) prostheses, and the walking condition were observed after surgery. The follow-up lasted from 6 to 25 months, with an average of 12.7 months.Results:The wound healing in 5 patients was good. The patients stood on the foot in 2 months after surgery, started to walk with the assistance of crutch in 3 months after surgery, and took walk without assistance in 5-6 months after surgery. Good osseous integration were achieved. One diabetic patient had infection of foot wound 3 months after surgery. After removal of microporous titanium prosthesis and replacement of vancomycin blended interstitial substance of bone cement, the wound healed and the patient resumed walking.Conclusion:It is an effective method to encourage the patients to take early ambulation after the surgery for lower extremity soft tissue defect with large bone defect that was repaired by a flap and 3D printed microporous titanium(tantalum)prosthesis. Further observations are required to investigate the long-term efficacy, and the reduction of prosthesis infection rate requires further exploration.