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

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.

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

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

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.

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.

Objective To investigate the effect and mechanism of mifepristone on the migration of human endometrial carcinoma cells. Methods A human endometrial carcinoma cell line, Ishikawa cells, was cultured in vitro and treated with mifepristone at different concentrations. Wound healing assay was applied to detect the migration of Ishikawa cells. RT?PCR and methylation?specific PCR (MSP) were used to detect the levels of H19 mRNA and its DNA methylation. Western?blot was used to detect the expressions of HMGA2 and epithelial to mesenchymal transition ( EMT ) related proteins. Results When treated with different concentrations of mifepristone for 48 hours, the width of scratch of the the control group, the 5 mg/L and the 10 mg/L mifepristone treatment groups were (4.18±0.07)mm,(4.68±0.07)mm, and(4.99±0.07) mm, respectively (P<0.05 for all) and treated with mifepristone for 72 hours, those were(3.46±0.07)mm, (4.29±0.07)mm, and(4.78±0.04)mm, respectively (P<0.05 for all). In the Ishikawa cells, mifepristone suppressed the transcriptional level of H19 through enhancing its promoter methylation, which resulted in inhibited expressions of HMGA2 and vimentin and increased expression of E?cadherin in a time? and concentration?dependent manner. Conclusion Mifepristone inhibits the migration of endometrial carcinoma cells partially through methylation?induced of transcriptional inhibition of H19, which results in the down?regulation of HMGA2 and vimentin and upregulation of E?cadherin.

Objective To investigate the effect and mechanism of mifepristone on the migration of human endometrial carcinoma cells. Methods A human endometrial carcinoma cell line, Ishikawa cells, was cultured in vitro and treated with mifepristone at different concentrations. Wound healing assay was applied to detect the migration of Ishikawa cells. RT?PCR and methylation?specific PCR (MSP) were used to detect the levels of H19 mRNA and its DNA methylation. Western?blot was used to detect the expressions of HMGA2 and epithelial to mesenchymal transition ( EMT ) related proteins. Results When treated with different concentrations of mifepristone for 48 hours, the width of scratch of the the control group, the 5 mg/L and the 10 mg/L mifepristone treatment groups were (4.18±0.07)mm,(4.68±0.07)mm, and(4.99±0.07) mm, respectively (P<0.05 for all) and treated with mifepristone for 72 hours, those were(3.46±0.07)mm, (4.29±0.07)mm, and(4.78±0.04)mm, respectively (P<0.05 for all). In the Ishikawa cells, mifepristone suppressed the transcriptional level of H19 through enhancing its promoter methylation, which resulted in inhibited expressions of HMGA2 and vimentin and increased expression of E?cadherin in a time? and concentration?dependent manner. Conclusion Mifepristone inhibits the migration of endometrial carcinoma cells partially through methylation?induced of transcriptional inhibition of H19, which results in the down?regulation of HMGA2 and vimentin and upregulation of E?cadherin.

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

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.