Objective : To explore the role of lysophosphatidylcholine acyltransferase 3(LPCAT3) in Erastin-induced ferroptosis of acute myeloid leukemia(AML) cells and its related molecular regulatory mechanisms. Methods : Tetrazolium salt(MTS) method was used to detect the sensitivity of different AML cells to the classic ferroptosis inducer Erastin, real time quantitative polymerase chain reaction(qPCR) was used to detect the basal expression level ofLPCAT3mRNA, and the correlation between them was analyzed. Lentivirus-mediatedLPCAT3overexpression AML cell lines(OE group) and negative control lines(NC group) were constructed. After Erastin intervention, MTS, flow cytometry, and micromethods were used to detect cell viability, lipid reactive oxygen species(ROS), and Malondialdehyde(MDA), respectively. qPCR and Western blot were used to detect unfolded protein response(UPR) classic pathway signaling molecules(PERK, ATF4, GRP78, etc.) expression levels. The above ferroptosis-related indicators were detected after combined intervention with the UPR inhibitor 4-phenylbutyric acid(4-PBA), and the regulatory relationship was analyzed. Results : Four different types of AML cells had different sensitivities to ferroptosis, among which K562 cells were relatively insensitive. The IC50of the four types of AML cells to Erastin was negatively correlated with the expression level ofLPCAT3(r=-0.919,P<0.001). After Erastin intervention, the cell viability of K562 cells in the OE group was significantly inhibited by Erastin compared with the NC group(P<0.001), and the levels of lipid ROS and MDA increased(P<0.001). The results of qPCR and Western blot showed that, compared with the NC group, the mRNA and protein expression of UPR classic pathway moleculesPERK,ATF4, andGRP78mRNA and protein increased in the OE group(P<0.01). After inhibiting the UPR pathway by 4-PBA, the viability of K562 cells decreased(P<0.01), and lipid ROS and MDA levels increased(P<0.01) compared with the uninhibited state. Conclusion Overexpression ofLPCAT3can promote ferroptosis in K562 cells, and this process is negatively regulated by the classical UPR pathway PERK/ATF.

Objective:To investigate the clinicopathological features of fumarate hydratase (FH) deficiency uterine leiomyoma.Methods:The data of 38 patients with FH deficiency uterine leiomyoma were screened and analyzed. The expressions of FH, S-(2-succino)-cysteine (2SC), desmin, p16, p53, CD 10 and cell proliferation associated nuclear antigen (Ki-67) proteins were detected by immunohistochemistry, and their clinicopathological features were analyzed retrospectively. Results:(1) Clinical features: the median age of the patients was (42.5±7.4) years old. Twenty-one cases (55%) of them were myomas found in physical examination, and the median maximum diameter of the tumor was 6.0 cm (range: 5.0-7.5 cm); myomectomy was performed in 23 cases (61%), total hysterectomy with or without bilateral appendages in 15 cases (39%); laparoscopic surgery in 27 cases (71%), open surgery in 11 cases (29%); none of the patients had renal cell carcinoma. (2) Histological features: atypical nuclear cells were distributed locally or diffusely, eosinophilic nucleoli and intranuclear inclusion bodies could be seen, glass like globules could be seen in the cytoplasm, nuclear division was 0-4/10 high power field (HPF), and antler like blood vessels and pulmonary edema-like changes could be seen in the stroma. Among 38 patients with FH deficiency uterine leiomyoma, FH was negative in 37 cases (97%), and positive in 1 case (3%); 2SC, desmin, p16, p53, CD 10 and Ki-67 showed focal positive expression in 38 cases (100%), including 35 cases (92%) with Ki-67 index<10% and 3 cases (8%) with Ki-67 index ≥10%. (3) Follow-up: 4 cases (11%) recurred, and there was no death. There were significant differences in age, family history, distribution of atypical nuclei and mitosis number between recurrent group and non-recurrent group (all P<0.05). Conclusions:FH deficiency uterine leiomyoma is a rare tumor, which needs pathological examination,immunohistochemical examination and clinical history. Patients younger than 43 years old, with family history, histologically atypical diffuse nuclear distribution and mitotic number ≥3/10 HPF should be alert to the risk of recurrence.

Thoracic aortic disease, which involves some of the arch vessels, is challenging to treat because of its complex anatomical structure and variation. With the progress and development of endovascular repair technology, in situ fenestration and pre-fenestration have become important measures for the treatment of thoracic aortic diseases. In recent years, there have been a series of reports on the application of in situ fenestration or pre-fenestration in a few centers, and the preliminary results are satisfactory. However, there are some problems such as single operation and small sample size. In order to further compare and analyze the safety and efficacy between the two operations, meta-analysis was conducted by searching Pubmed, Embase, Cochrane, Web of Science, China National Knowledge Infrastructure, Wanfang data, VIP data and China Biology Medicine disc. There was no significant difference in technical success rate, 30-day mortality, endoleak rate and reintervention rate between the two methods. The application of surgical methods can be determined according to the patient's condition and the surgeon's clinical experience.

Objective To investigate the effect of piR-9994 on the biological behavior of gastric cancer cells and its possible mechanism. Methods The expression of piR-9994 in gastric cancer cell lines (MGC803 and AGS) and normal gastric epithelial cells (GES-1) were detected by qRT-PCR. MGC803 cell line with piR-9994 overexpression and knockdown were constructed. The effects of piR-9994 expression changes on cell proliferation were detected by MTT and clone formation assay. The scratch wound healing assay and Transwell invasion assay were used to detect cell migration and invasion abilities. qRT-PCR and Western blot were used to detect cell proliferation and EMT-related genes expression. Results The expression level of piR-9994 in MGC803 cells was significantly higher than that in normal gastric epithelial cell line GES-1 (P < 0.05). The overexpression of piR-9994 promoted the proliferation, invasion and metastasis of gastric cancer cells, while piR-9994 knockdown had the opposite effect. piR-9994 expression was closely related to cell cycle, especially in S phase. The overexpression of piR-9994 promoted the expression of proliferation-related genes PCNA, CCND1 and Bcl-2, inhibited the expression of apoptosis gene C-PARP, and promoted the expression of EMT-related genes N-cadherin, MMP7, Twist and Vimentin; while piR-9994 knockdown had the opposite effect. Conclusion Abnormal expression of piR-9994 affects the proliferation, invasion, metastasis and EMT process of gastric cancer cells. piR-9994 may be a new biomarker and therapeutic target for gastric cancer.

Objective To explore the expression of circ_0006692 in NSCLC and its relation with clinicopathological characteristics and related mechanism. Methods We collected 50 pairs of NSCLC tissues and adjacent tissues. The expression of circ_0006692 was detected by qRT-PCR and its relation with clinicopathological features was analyzed. We constructed lung cancer A549 cell line with circ_0006692 overexpression and knockdown. Cell proliferation, migration and invasion were detected by MTS, colony forming, wound healing and Transwell invasion assays. qRT-PCR and Western blot were used to detect the effect of circ_0006692 expression change on EMT-related gene expression. Results The expression of circ_0006692 in NSCLC was significantly higher than that in adjacent tissues (P < 0.05). The expression level of circ_0006692 was closely related to tumor size, TNM stage and pulmonary membrane invasion (P < 0.05). circ_0006692 knockdown inhibited cell proliferation, invasion and metastasis, while its overexpression promoted cell proliferation, invasion and metastasis. circ_0006692 knockdown inhibited the expression of EMT-related genes CDH2 and MMP7 in A549 cells and promoted the expression of CDH1. Conclusion The upregulated expression of circ_0006692 in NSCLC is closely related to tumor size, TNM stage and pulmonary membrane invasion. circ_0006692 expression could regulate the proliferation, invasion, metastasis and EMT progression of lung cancer A549 cells.

Objective To explore the efficacy of temporary titanium plate screwing for positional maintenance in reduction and internal fixation for displaced acetabular fractures.Methods A retrospective study was conducted of the 28 patients (28 hips) with displaced acetabular fracture who had been treated by open reduction and internal fixation from October 2013 to March 2016.They were 20 males and 8 females,aged from 24 to 68 years (average,42.3 years).The time from injury to surgery ranged from 7 to 21 days(average,14.5 days).According to the Letournel-Judet classification,there were 2 posterior column fractures,4 transverse fractures,5 posterior column plus posterior wall fractures,6 transverse plus posterior wall fractures,2 T-shaped fractures,3 anterior and posterior transverse fractures and 6 double column fractures.The posterior acetabular approach or combined anterior and posterior approach was used.In all the patients temporary titanium plate screwing was conducted to fix one side of the fracture so as to facilitate accomplishment of open reduction and internal fixation without losing the indirect anatomic reduction of the intraarticular fracture.After the open reduction and internal fixation was accomplished,the temporary screw fixation was removed in 26 patients but retained in 2 patients as needed.The reduction quality,complications and outcomes at the final follow-ups were recorded.Results All the 28 patients were followed up for 10 to 36 months (mean,15.6 months).By the Matta criteria,anatomical reduction was achieved in 26 cases,unsatisfactory reduction in one and poor reduction in one,yielding an anatomic reduction rate of 92.9％.By the improved Mere d'Aubigne & Postel criteria,the clinical outcomes at the final follow-up were excellent in 27 cases and good in one,yielding a good to excellent rate of 100％.Postoperatively,heterotopic ossification of different severities occurred in 11 cases but did not affect their joint function;transient paralysis of the sciatic nerve was reported in 6 cases but recovered 3 months after surgery.No complications like avascular necrosis of the femoral head or walking pain were observed during follow-ups.Conclusion Temporary fixation with titanium plate screws during open reduction and internal fixation for displaced acetabular fractures can effectively improve the reduction and fixation of the articular surface,leading to satisfactory short-term clinical outcomes.

Objective To investigate the expressiorn of microRNA-622(miR-622) and dual specificity tyrosine phosphorylation-regulated kinase 2 (DYRK2) in colon cancer tissues and cell lines and explore the effect of miR-622 on SW11l6 cells migration and invasion.Methods Eighty-two colon cancer tissues and paired para-tumor tissue specimens were collected.C.olon cancer cell line SW1116,SW480 and normal human colon epithelial cell line NCM460 were cultured.MiR-622 was detected by using Real time PCR,DYRK2 expression was measured by using immunohistochemistry,Real time PCR anid Western blot in tissue level and cell level,respectively.The relation of miR-622 and DYRK2 was analyzed by Pearson correlation analysis.miR-622 mimics transfection was conducted to up-regulate miR-622,while negative control,NC group were transfected with control sequence.Expression of DYRK2 was evaluated by using Real time PCR and Western blot,while Transwell chamber assays were used to assess the migration ability changes.Results Real time PCR and Western blot results showed that miR-622 mRNA was highly expressed in colorectal cancer tissue and colon cancer cell SW1116,whereas DYRK2 mRNA and protein were lowly expressed when compared with paracancerous tissue and normal colonic epithelial cell line NCM460.An obvious negative correlation was showed between miR-622 and DYRK2(r=0.916,P＜0.01).Compared to NC group,DYRK2 mRNA and protein expression were down-regulated after transfection of miR-622 mimics,which was observerd through Real time PCR and Western blot(P＜0.01).Correspondingly,compared to NC group,the migration ability of SW116 was remarkably enhanced after transfection of miR-622 mimics(P＜0.01).Conclusion The expression of miR-622 is high and DYRK2 is low in colon cancer.Up-regulation of miR-622 could negatively regulate DYRK2 expression and promote SW1116 cells migration.

Objective: To investigate the expression of long non-coding RNA (lncRNA) SNHG8 in EB virus related gastric cancer and their correlation prognosis. Methods: The expression of SNHG8 in 93 gastric cancers and 93 cancer-free controls, matched by age and sex, were determined by real-time PCR. EB virus expression was detected by EBER in situ hybridization. Results: Forty-one gastric cancers were EB virus associated. For all gastric cancers, SNHG8 expression was 14 times higher (P=0.001) than that in non-cancer controls; in the EB virus related gastric cancers, SNHG8 expression was increased 25 times (P<0.05) over EB virus negative gastric cancers. SNHG8 expression level was also significantly associated with TNM staging (P<0.05). Conclusions SNHG8 may act as a proto-oncogene, participating in gastric carcinogenesis.EB virus infection of gastric mucosa may promote SNHG8 expression.

Objective: To investigate the clinicopathologic features of microcystic, elongated and fragmented (MELF) pattern invasion of endometrial adenocarcinoma. Methods: HE and immunohistochemistry staining method were used to analysis morphologic features and immunophenotype of 72 patients of endometrial adenocarcinoma with MELF pattern invasion, and chi-square test was used to analysis the clinicopathologic features. Results: The mean age of 72 patients was 54 years (40 to 70 years). Thirty-two patients were pre-menopausal and 40 were post-menopausal. According to the FIGO staging system (2014), 32 cases(44.4%)were at stage Ⅰ, 22 cases(30.6%)at stage Ⅱ, 17 cases(23.6%)at stage Ⅲ and 1 case(1.4%) at stage Ⅳ. Microscopically, MELF invasion showed microcystic, elongated slit-like or fragmented glands in myometrium and their lining cells usually were cube or flat, as well as the single or clusters of eosinophilic tumor cells mimicking histocytes. In addition, a fibromyxoid or inflammatory stromal response was often present.Immunohistochemical staining showed that MELF invasion was positive for p16, CA125 and CA19-9, but negative for ER, PR and p53.Compared with non-MELF pattern invasion, significant differences were noted in menopause pausimenia, FIGO stages, deep invasion into myometrium, lymph metastasis, lymphovascular space invasion (LVSL), serum CA125 and CA19-9 in patients with MELF pattern invasion (all P<0.05). Conclusions MELF pattern invasion of endometrial adenocarcinoma is characterized by advanced FIGO stage, deep myoinvasion, high metastasis rate to lymph node and LVSL. Pathologists should recognize the MELF invasion and evaluate the depth of myometrium of infiltration and LVSL with special attention to the presence of MELF invasion with necessary immunohistochemistry for more accurate pathological diagnosis.

OBJECTIVETo investigate the methylation status of Runx3 promoter and Runx3 expression in breast lesion tissues.

METHODSOne hundred and fourteen breast lesions, including 35 cases of fibroadenoma, 39 cases of intraductal carcinoma, 40 cases of invasive ductal carcinoma, and 33 cases of normal breast tissue from Fabruary 2010 to August 2012 were included in this study. Runx3 protein expression was assessed by immunohistochemical SP method; whereas methylation of Runx3 promoter was assessed by high resolution melting (HRM) analysis.

RESULTSRunx3 protein was mainly expressed in the cytoplasm of ductal epithelial cells. The expression rates of Runx3 in normal breast tissue, fibroadenoma, ductal carcinoma in situ, invasive ductal carcinoma were 87.9% (29/33), 85.7% (30/35), 53.8% (21/39), and 40.0% (16/40) respectively. The methylation rates of Runx3 promoter were 12.1% (4/33), 20.0% (7/35), 46.2% (18/39), and 57.5% (23/40), respectively. Correlation analysis between promoter methylation and protein expression of Runx3 in different breast tissue showed the r value in normal breast tissue, fibroadenoma, ductal carcinoma in situ and invasive ductal carcinoma was -0.431 (P = 0.012), -0.408 (P = 0.015), -0.589 (P = 0.000) and -0.743 (P = 0.000) respectively.

CONCLUSIONSRunx3 protein expression shows a downward trend in ductal carcinoma in situ and invasive ductal carcinoma, meanwhile its promoter methylation increases significantly. The methylation of Runx3 promoter may be one of the important factors in the occurrence and development of breast cancer.

Breast ; metabolism ; Breast Neoplasms ; metabolism ; Carcinoma, Ductal, Breast ; metabolism ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; DNA Methylation ; Female ; Fibroadenoma ; metabolism ; Humans ; Neoplasm Proteins ; metabolism ; Promoter Regions, Genetic