Potential of thrombospondin-1 in treatment of polycystic ovary syndrome rat model: a preliminary study.

OBJECTIVE: Polycystic ovary syndrome (PCOS) is a common gynecological endocrine disease in reproductive women, and the endocrine levels are also affected by diseases. The aim of this study was to determine the effect of thrombospondin-1 (TSP-1) on PCOS rat model. METHODS: We established the PCOS rat model, the serum hormones including TSP-1 expression were determined and morphological characteristics were investigated to evaluate the model. These above endocrine and morphological features were investigated again to evaluate the effect of TSP-1 treatment. RESULTS: In the PCOS model group, the serum hormones change (higher luteinizing hormone, testosterone and estrogen) and decreased TSP-1 expression levels were found compared with the control group. Besides, the morphological characteristics of PCOS were also observed in the model group. After TSP-1 treatment, the higher TSP-1, ANGPT2, PDGFB and PDGFD expression levels, the lower LH and T levels, decreased vessel density as well as VEGFA and ANGPT1 expression levels were found compared with the control group, and the ovary morphological changes were also observed in the TSP-1 experimental group. CONCLUSIONS: TSP-1 delivery system might be an alternative therapy for PCOS treatment.

Impact of Platelet Counts, Surgical Methods, and Preoperative Platelet Transfusion on the Outcome of Splenectomy for Immune Thrombocytopenia.

INTRODUCTION: Splenectomy is an important and potentially curative treatment for immune thrombocytopenia (ITP). Laparoscopic splenectomy (LS) has replaced open splenectomy (OS) as the standard approach. The prognostic role of platelet count and the clinical indication of preoperative platelet transfusion are not entirely clear. METHODS: We designed a study to explore the prognostic impact of surgical methods, platelet count, and platelet transfusion in a large, single-institute, long-term cohort of ITP patients. RESULT: In 118 ITP patients, there was no difference between OS and LS in response and surgical complications. The overall response rate was 77% and the complete response (CR) rate was 70%. Patients with a CR had a trend towards a higher baseline platelet count. A stable platelet count 14-28 days after splenectomy was associated with a sustained long-term response. Patients requiring preoperative platelet transfusion had a lower preoperative platelet count and were more likely to need postoperative transfusion of red blood cells and platelets. They also had a lower postoperative platelet count than the nontransfusion group. Relapse-free survival did not differ. CONCLUSIONS: Baseline and postoperative platelet counts are apparently associated with the treatment response to splenectomy but the difference did not reach statistical significance. Preoperative platelet transfusion did not overcome the disadvantage of thrombocytopenia and was not recommended when other preparative measures are available.

A model based on CT radiomic features for predicting RT-PCR becoming negative in coronavirus disease 2019 (COVID-19) patients.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has emerged as a global pandemic. According to the diagnosis and treatment guidelines of China, negative reverse transcription-polymerase chain reaction (RT-PCR) is the key criterion for discharging COVID-19 patients. However, repeated RT-PCR tests lead to medical waste and prolonged hospital stays for COVID-19 patients during the recovery period. Our purpose is to assess a model based on chest computed tomography (CT) radiomic features and clinical characteristics to predict RT-PCR negativity during clinical treatment. METHODS: From February 10 to March 10, 2020, 203 mild COVID-19 patients in Fangcang Shelter Hospital were retrospectively included (training: n = 141; testing: n = 62), and clinical characteristics were collected. Lung abnormalities on chest CT images were segmented with a deep learning algorithm. CT quantitative features and radiomic features were automatically extracted. Clinical characteristics and CT quantitative features were compared between RT-PCR-negative and RT-PCR-positive groups. Univariate logistic regression and Spearman correlation analyses identified the strongest features associated with RT-PCR negativity, and a multivariate logistic regression model was established. The diagnostic performance was evaluated for both cohorts. RESULTS: The RT-PCR-negative group had a longer time interval from symptom onset to CT exams than the RT-PCR-positive group (median 23 vs. 16 days, p < 0.001). There was no significant difference in the other clinical characteristics or CT quantitative features. In addition to the time interval from symptom onset to CT exams, nine CT radiomic features were selected for the model. ROC curve analysis revealed AUCs of 0.811 and 0.812 for differentiating the RT-PCR-negative group, with sensitivity/specificity of 0.765/0.625 and 0.784/0.600 in the training and testing datasets, respectively. CONCLUSION: The model combining CT radiomic features and clinical data helped predict RT-PCR negativity during clinical treatment, indicating the proper time for RT-PCR retesting.

Immune thrombocytopenia: Effectiveness of frontline steroids and comparison of azathioprine, splenectomy, and rituximab as second-line treatment.

OBJECTIVE: For immune thrombocytopenia (ITP), efficacy of frontline steroids is well established. However, clinical data comparing various treatment options for refractory or relapsed ITP are limited. We aimed to investigate the outcome of frontline steroid treatment for ITP patients and compare common second-line modalities in a single institute in Taiwan. METHODS: We collected the complete outpatient list over a 6-month period. Patients were identified from the list, and medical records were reviewed to capture the data retrospectively. The diagnosis of ITP was made by excluding other etiologies. RESULTS: Among 665 patients with thrombocytopenia, the diagnosis of ITP was made in 375. Two hundred and fifty-seven patients (51 males, median age 45.5) received treatment. Response to steroids was evaluable for 184 patients. Complete response (CR) was achieved in 120 (65.2%) and partial response in 43 (23.3%). In 21 (11.4%) patients, ITP was refractory to steroids. Among those with CR, 76 (63%) patients relapsed in a median of 9.5 months. After relapse or steroid failure, 57 (49%) received azathioprine treatment and 38 (32%) underwent splenectomy. Response rate was 71.4% (38.1% CR) for azathioprine and 91.4% (77.1% CR) for splenectomy. Rituximab was effective in 8 of 10 patients. CONCLUSION: Steroids are effective frontline treatment for ITP, but relapse is common. Both azathioprine and splenectomy are effective treatment after steroid failure. Rituximab appears to a reasonable second-line treatment option in our limited experience.

Sleep Deprivation and Late Bedtime Impair Sperm Health Through Increasing Antisperm Antibody Production: A Prospective Study of 981 Healthy Men.

BACKGROUND The aim of this study was to investigate the effects of sleep duration and bedtime on sperm health, and the possible mechanism involved. MATERIAL AND METHODS We randomly divided 981 healthy Chinese men into groups according to research-set bedtimes (A=8-10 PM, B=after 10 PM, and C=after midnight) and sleep durations: group 1=<6.0 h (short), group 2=7.0-8.0 h (average), and group 3=>9.0 h (long). Sperm morphology, count, survival, and motility were examined according to sleep patterns. Antisperm antibody (ASA) production in semen was determined. RESULTS Sperm counts and their survival rates were lower in the short sleepers as compared to others within each group (all P<0.01). The lower counts and survival rates were observed in different bedtimes, with significant differences found between measurements of C1 vs. A1 and C2 vs. A2 or B2 (all P<0.05 or 0.01). Semen motility was lower in the short sleepers as compared to the average and long sleepers (all P<0.01). There were differences in the bedtime-related results between measurements of C1 vs. A1 or B1 (P<0.05 or 0.01). Additionally, the population proportion for the ASA-positive participates and incidence of the ASA-expressed population obviously increased in the short sleepers as compared to others within each group (all P<0.05). CONCLUSIONS Short and long sleep durations and late bedtime were associated with impaired sperm health in the study cohort, partly through increasing ASA production in the semen.

5-Hydroxymethylcytosine expression is associated with poor survival in cervical squamous cell carcinoma.

OBJECTIVE: Deoxyribonucleic acid methylation is an important epigenetic modification that is frequently altered in cancer. Recent reports showed that the level of 5-hydroxymethylcytosine was altered in various types of cancers. The influence of deoxyribonucleic acid methylation in cervical squamous cell carcinoma is not fully understood. In this study, we investigated 5-hydroxymethylcytosine and ten-eleven translocation expression in cervical squamous cell carcinoma and whether they are associated with poor survival in cervical squamous cell carcinoma. METHODS: We detected the expression of 5-hydroxymethylcytosine, 5-methylcytosine and TET1/2/3 in 140 patients with cervical squamous cell carcinoma and 40 patients with normal cervical tissues by immunohistochemistry. We assessed the prognostic values of 5-hydroxymethylcytosine, 5-methylcytosine and TET2 in the clinical outcome of cervical squamous cell carcinoma. RESULTS: Expression of 5-hydroxymethylcytosine was significantly decreased in cervical squamous cell carcinoma compared with normal cervix tissues. In contrast, 5-methylcytosine expression was significantly increased in cervical squamous cell carcinoma compared with normal cervix tissues. Moreover, expression of TET2, but not TET1 and TET3, was decreased in cervical squamous cell carcinoma. Our study showed that the decreased level of 5-hydroxymethylcytosine predicts poor prognosis of cervical squamous cell carcinoma patients. The expression of 5-hydroxymethylcytosine was an independent prognostic factor for both disease-free and overall survival of cervical squamous cell carcinoma patients. CONCLUSIONS: In cervical squamous cell carcinoma, less aggressive tumor behavior was correlated with 5-hydroxymethylcytosine and TET2. Our data indicated that 5-hydroxymethylcytosine may become a prognostic marker for cervical squamous cell carcinoma and the decreased expression of TET2 may be an underlying mechanism for decreased 5-hmC in cervical squamous cell carcinoma.

Prognostic values of 5-hmC, 5-mC and TET2 in epithelial ovarian cancer.

PURPOSE: DNA methylation is an important epigenetic modification that is frequently altered in cancer. Recent reports showed that the level of 5-hydroxymethylcytosine (5-hmC) was altered in various types of cancers. The influence of DNA methylation in epithelial ovarian cancer (EOC) is not fully understood. Therefore, the aim of the present study was to investigate factors involved in DNA demethylation in EOC compared with normal ovarian tissues. METHODS: We examined the expression of 5-hmC, 5-mC, and TET2 by immunohistochemistry in 130 cases of EOC and 40 cases of normal ovarian tissues. We assessed the prognostic values of 5-hmC, 5-mC, and TET2 in clinical outcome of EOC. RESULTS: We discovered a significant decrease in 5-hmC and TET2 expression in EOC compared with normal ovarian tissues. In contrast, there was a significant increase in 5-mC expression in EOC compared with normal ovarian tissues. The expression of 5-hmC, 5-mC, and TET2 correlated with pathologic stage, tumor grading, lymph node metastasis, and vascular thrombosis. Furthermore, decreased level of 5-hmC predicts poor prognosis of EOC patients. The expression of 5-hmC was an independent prognostic factor for overall survival of EOC patients. CONCLUSIONS: The data suggest that loss of 5-hmC is an epigenetic event of EOC, and the expression of 5-hmC could serve as a prognostic factor for EOC.

'Struggling to be an insider': a phenomenological design of new nurses' transition.

AIMS AND OBJECTIVES: To obtain a comprehensive understanding of the transition process of new nurses in Taiwan. Background. The transition period for new nurses can be a daunting and traumatic experience. The large number of newly graduated Taiwanese nurses who resign from their jobs within three months indicates that this process can be complicated. However, the problems associated with the experiences of new nurses have not yet been recognised. DESIGN: We adopted a phenomenological design using focus group interviews. METHODS: Sixteen new nurses (less than one year working experience) participated in eight weekly group interviews lasting two hours each to grasp their experience of 'being new'. Interview data were analysed according to Sloan's (2002) three moments, and the whole process of analysis followed the suggestions of Agar (1986), which was performed in a close collaboration between researchers until the consensus about the findings could be reached. RESULTS: The overarching pattern of the transition process of new nurses becoming experienced members of the clinical nursing team was revealed as a journey of 'struggling to be an insider'. This phenomenon was characterised by four themes, including (1) 'being new as being weak', (2) 'masking myself', (3) 'internalising the unreasonable' and (4) 'transforming myself to get a position'. CONCLUSIONS: While Western culture view abusive indoctrination of new nurses as toxic behaviour, under the Chinese traditions of yield, tolerance and self-oppression, following the power hierarchy and seeking harmony, the transition of new nurses is interpreted differently. RELEVANCE TO CLINICAL PRACTICE: Recognition of the journey of 'struggling to be an insider' helps nurse administrators to (1) gain a better understanding of what new nurses encounter in their transition process, (2) help new nurses without harm, (3) improve in-service training programmes and (4) retain future nurses.

Relationships between the coronary fat attenuation index for patients with heart-related disease measured automatically on coronary computed tomography angiography and coronary adverse events and degree of coronary stenosis.

Background: Pericoronary artery coronary tissue (PACT) is a type of epicardial fat that can reflect the state of the coronary artery (inflammation, etc.). However, it cannot be reasonably and efficiently utilized in routine computed tomography (CT) examination. The aim of this study was to use artificial intelligence (AI) software to analyze coronary computed tomography angiography (CCTA) and measure the coronary perivascular fat attenuation index (FAI) of patients. The relationship between FAI and the occurrence of coronary adverse events and the degree of coronary stenosis were further analyzed. Methods: This study involved patients who experienced CCTA in West China Hospital, Sichuan University, from January 2012 to December 2012. These patients were followed up to 2020 and classified according to the occurrence of coronary adverse events and the degree of stenosis of the lumen. For all patients, AI software was used to analyze the CCTA images of patients, and the FAI of 3 coronary arteries, the left anterior descending artery (LAD), the left circumflex artery (LCX), and the right coronary artery (RCA), was measured. Moreover, the relationship between FAI and patients with different degrees of coronary stenosis and adverse coronary events was determined. Results: Comparisons between any 2 groups showed that the differences in the FAI among the 4 groups for the LAD were significant (all P values <0.05). There were no significant differences between the group with less-than-moderate stenosis (Mb) without adverse events and the group with moderate-or-above stenosis (M) with no adverse events for the LCX (P>0.05). For the remaining groups, FAI values exhibited statistically significant differences (P<0.05). According to the degree of lumen stenosis, the patients were divided into groups according to LAD, LCX, and RCA and the sum of the 3 vessels. There were significant differences in coronary FAI among the groups with different degrees of lumen stenosis for the sum of the 3 vessels, the LAD, and the LCX (P<0.05). Conclusions: FAI can reflect the state of the coronary artery, which is related to inflammation of the coronary lumen. Moreover, there is a relationship between FAI and the degree of stenosis in the coronary lumen: the narrower the coronary lumen is, the higher the FAI around the lumen.

[Clinical value of anti-adhesion agents used in laparotomy in obstetrics and gynecology].

OBJECTIVE: To investigate clinical value of anti-adhesion agent:chitosan in preventing adhesions followed by laparotomy surgery in obstetrics and gynecology. METHODS: From Jan. 2006 to Dec. 2009, 770 patients underwent laparotomy surgery at Department of Obstetrics and Gynecology in the Second Affiliated Hospital of Harbin Medical University. One hundred and twenty-five patients underwent secondary surgery due to disease recurrence or cesarean section, the previous surgery were 18 cases with myomectomy, 20 cases with endometriosis surgery, 5 cases with resection of pelvic abscess, 20 cases with ectopic pregnancy surgery, 27 cases with benign adnexal neoplasm resection, 9 cases with cyto-reductive surgery in ovarian cancer, 26 cases with variable infertility surgery. Those 125 patients were managed by different agents washing before abdomen closure, which were assigned into two groups, including 59 cases washed by saline in control group and 66 cases washed by chitosan in study group. The abdomen adhesion in secondary surgery was evaluated by adhesion classification system. RESULTS: In study group, 37 patients without adhesions, 20 patients with degree I adhesion, 6 patients with degree II adhesion, 3 patients with degree III adhesion, and no patients with degree IV adhesion were observed. While in control group, there were 11 patients without adhesions, 23 patients with degree I adhesion, 14 patients with degree II adhesion, 8 patients with degree III adhesion, and 3 patients with degree IV adhesion. The distribution of adhesion reached statistical significance between the two groups (χ(2) = 20.9999, P = 0.0003). Twenty-six patients in previous surgeries due to infertility included 17 cases in control group and 9 cases in study group. They all were managed by cesarean section in secondary surgery, it was found that 15 cases without adhesion and 2 cases with degree I adhesion were in study group and 2 cases without adhesion and 7 cases with degree I adhesion were in control group. It reached statistical difference (P = 0.0016). CONCLUSION: Anti-adhesion agent could prevent adhesion followed by surgery in obstetrics and gynecology effectively.

[Value of coronary CT angiography in assessment of bifurcation lesions].

OBJECTIVE: To evaluate the value of coronary CT angiography in assessment of bifurcation lesions. METHODS: The original image of 79 established and suspected coronary artery disease patients who underwent both coronary CT angiography and conventional artery angiography (CAG) sequentially were included in this analysis. Bifurcation lesions were assessed on primary and secondary vessels with diameter ≥ 2.0 mm, bifurcation lesions were graded according to Chen's classification. CAG was used as golden standard. The sensitivity, specificity, positive predictive value and negative predictive value were calculated. Spearman's test and Kappa test were used to evaluate the correlation and classification identity of the two methods. RESULTS: CAG evidenced 177 bifurcation lesions out of 445 bifurcation vessels and coronary CT detected 168 bifurcation lesions out of 404 bifurcation vessels with satisfactory imaging quality and 390 bifurcation vessels could be analyzed by both CAG and coronary CT. Sensitivity, specificity, positive predictive value and negative predictive value of coronary CT angiography were 94.2%, 94.6%, 90.7%, 96.1%, respectively. The results for the lesions at LM-LAD/LCX + LAD/Mid, LAD/Diag, RCA/PDA were more satisfactory and the sensitivity and specificity were as high as: 97.1% and 94.2%, 95.7% and 89.5%, 92.3% and 98.7%, respectively. There were significant correlations for evaluating the narrow degree of the opening of the bifurcation branch with these two methods (r = 0.799 58, P < 0.01) and for identifying I, II, III type bifurcation lesions (Kappa coefficient = 0.7959, P < 0.01) as well as for identifying the subtype bifurcation lesions (Kappa coefficient = 0.6328, P < 0.01) using the two methods. CONCLUSION: Coronary CT angiography is efficient in identifying the bifurcation lesions and offers a reasonable indication for bifurcation lesion classification.

Changing profile of platelet activity and turnover indices during treatment response of immune thrombocytopenia.

Both platelet count and function change after treatment of immune thrombocytopenia. Platelet function can be measured by plasma markers, including platelet activity [e.g., soluble P-selectin (sP-selectin) and soluble CD40 ligand (sCD40L)] and platelet turnover markers [e.g., glycocalicin (GC)]. Patients were classified into no response (NR, including new diagnosis), partial response (PR) and complete response (CR). One hundred and sixteen samples (29 CR, 32 PR, 55 NR) from 79 patients were collected. Plasma markers (sP-selectin, sCD40L and GC) were measured by ELISA. Platelet counts and mean platelet volume (MPV) were obtained in the clinical laboratory using GenS System-2. The results showed that responsive patients (PR + CR) had higher levels of sP-selectin (P = 0.026) and sCD40L (P = 0.001). Although there was no difference in MPV (P = 0.077) or GC (P = 0.078), there was a marked decrease of GC index (P < 0.001) in responsive patients. Paired sample analysis showed no difference in sP-selectin, sCD40L, MPV or GC but significant difference in GC index (P = 0.017) between NR and PR. Another paired sample analysis showed no difference in sP-selectin, sCD40L, MPV or GC but significant difference in GC index (P = 0.029) between PR and CR. Patients with refractory and newly diagnosed disease had a significant difference in GC (P = 0.020) and sCD40L (P = 0.001), despite similarly low platelet counts. In conclusion, platelet activity markers (sP-selectin and sCD40L) and GC indices change in parallel with treatment response. Plasma levels of GC and sCD40L may be predictors of treatment response.

Variations in the Profiles of Vascular-Related Factors Among Different Sub-Types of Polycystic Ovarian Syndrome in Northern China.

Recently, a growing body of evidence has suggested that abnormal ovarian angiogenesis, secondary to the imbalance between various angiogenic markers, is involved in the pathogenesis of PCOS, and this has led to the use of various interventions (such as Diane-35) to restore the normal ovarian angiogenesis. Therefore, we conducted the current investigation to determine the role of such markers (endothelial growth factor (VEGF), endostatin (ES), and thrombospondin-1 (TSP-1)) in the pathogenesis of PCOS along with the associated changes in ovarian blood flow in patients with PCOS compared to healthy controls, both before and after a course of oral contraception. A total of 381 patients with PCOS and 98 healthy females of childbearing age were recruited from July 2014 to June 2017 at the Reproductive Center of the Second Affiliated Hospital of Harbin Medical University. The serum levels of VEGF, ES, and TSP-1 were determined by enzyme-linked immunosorbent assay, while ovarian perfusion was measured by the pulsatility index (PI) and resistance index (RI) by using transvaginal color Doppler ultrasound. Repeated analyses were carried out after 3 months of Diane-35 treatment. Post-treatment serum levels of luteinizing hormone (LH)/follicle stimulating hormone (FSH) ratio of patients with PCOS decreased significantly (P <0.05). The RI values of most PCOS patients increased after treatment (P<0.05), while PI was significantly increased in all patients (P<0.05). However, variable changes in the serum levels of TSP-1, VEGF, and ES after treatment were observed. Serum VEGF levels showed a negative correlation with serum LH/FSH ratio, T concentration, and ES (P <0.05), while ES levels were negatively correlated with serum T concentrations only (P<0.05). The markers of angiogenesis (VEGF, ES, and TSP-1) were expressed differently among PCOS patients, who also responded differently to the same course of Diane-35 treatment. This field still warrants further investigation to reach a more definitive conclusion.

Primitive neuroectodermal tumors: a clinical and radiological analysis of six cases.

BACKGROUND: Primitive neuroectodermal tumor (PNET) is extremely rare and highly aggressive with poor prognosis. Few studies concerning PNET's the imaging features have been published. METHODS: Six cases of PNET, all confirmed by pathology and immunohistochemical (IHC) examinations, were treated during January 2012 to December 2016. These cases' clinical course and imaging findings were retrospectively analyzed. RESULTS: Among six PNET cases, one located in left superior abdomen, one case at posterior abdominal wall, one case in right orbit, one case in left frontal temporal lobe, one case in pelvic cavity, and one case located in left supraclavicular fossa. The tumor's density was uniform for small tumor and heterogeneous for large tumors on CT images, while the size of tumors differed during presentation depending on the location of the tumor. Marked enhancement was visualized after injection of contrast media. The demarcation between the lesion and adjacent tissues or organs tended to be unclear. On magnetic resonance imaging (MRI) images, the mass mainly showed heterogeneously long T1 and long T2 signal intensity, mixed high signal intensity on fluid-attenuated inversion recovery (FLAIR) image. In two cases maximum intensity projection image reconstruction demonstrates tortuous blood vessels within the tumor on enhanced CT images. Five cases were treated by surgical resection with two cases received adjuvant radiotherapy and three cases received adjuvant chemotherapy. Six patients were followed up with a mean period of 34.5 months (ranging from 6 to 55 months). Five patients survived and one died. Among the five patients undergoing surgeries, one patient presented pelvic and abdominal recurrence/metastasis 2 months after abdominal PNET resection. One patient had a recurrent lesion in the right orbit involving the right ethmoid sinus 6 months after right orbital PNET resection. One patient's pelvic tumor recurred 7 months after PNET operation, and this patient died after 1 year and 10 months of follow-up. During the follow-up period, the remaining three cases did not show obvious recurrence and/or metastasis. CONCLUSIONS: Overall, the imaging appearances of PNET lack characteristics. PNETs generally do not have clear boundary, or partially so, with its adjacent organs or tissues suggesting their invasive nature. Upon further validation, maximum intensity projection image reconstruction demonstrates tortuous blood vessels within the tumor on enhanced CT images may be valuable information for the diagnosis of PNET.

Long noncoding RNA SBF2-AS1 act as a ceRNA to modulate cell proliferation via binding with miR-188-5p in acute myeloid leukemia.

LncRNA SBF2-AS1 has been reported to be implicated in the deterioration of multiple human cancers. However, the roles and underlying mechanisms of SBF2-AS1 in acute myeloid leukemia (AML) are still unclear. In the present study, the online GEPIA database showed that SBF2-AS1 expression was significantly increased in AML samples. QRT-PCR results showed that SBF2-AS1 expression was upregulated in AML cells. CCK-8 assay revealed that SBF2-AS1 inhibition decreased AML cells proliferation ability in vitro. Flow cytometry assays showed that SBF2-AS1 inhibition induced AML cells apoptosis and arrested AML cells in G0/G1 phase. Mechanistically, miR-188-5p was identified as a direct target of SBF2-AS1. SBF2-AS1 upregulated the expression level of ZFP91 by sponging miR-188-5p. And the effects of SBF2-AS1 suppression on AML cells progression could be abolished by miR-188-5p inhibitors. Moreover, we found that SBF2-AS1 inhibition reduced tumor growth in vivo. Taken together, our findings elucidated that SBF2-AS1 could act as a miRNA sponge in AML progression, and provided a potential therapeutic strategy for AML treatment.

[Inhibitory effect of antisense oligonucleotide of integrin-linked kinase on cell proliferation of human epithelial ovarian cancer].

OBJECTIVE: To explore the inhibitory effect of integrin-linked kinase antisense oligonucleotide (ILK-ASODN) on cell proliferation in human ovarian cancer cell line (HO8910). METHODS: We transfected ILK-ASODN into HO8910 to block ILK gene expression, measured the expression levels of integrin-linked kinase (ILK) mRNA by RT-PCR and ILK protein by western-blotting; the inhibiting effects of the transfection on HO8910 proliferation, the cell cycles, and cell apoptosis were assessed by water soluble tetrazolium-1 (WST-1) and flow cytometry (FCM). RESULTS: After transfection of ILK-ASODN, the expression levels of ILK mRNA decreased significantly in groups D, E, F being 0.307 +/- 0.011, 0.198 +/- 0.008, 0, respectively, when compared with those of the two control groups of A and B (P < 0.05). The expression levels of ILK protein of the groups D, E and F decreased significantly also, being 26.3 +/- 0.8, 20.6 +/- 0.4 and 0, respectively. HO8910 cell proliferation was inhibited significantly, and the rates of apoptosis of the groups D, E and F increased significantly, being 7.31%, 8.84% and 11.27% respectively. The cell population increased in G0/G1 phase of the groups D, E and F, being 49.25%, 56.28% and 67.61% respectively, significantly different in comparison with those of groups of A and B (P < 0.01). CONCLUSIONS: Transfection of ILK-ASODN into human ovarian cancer line inhibited cancer cell proliferation significantly.

An Observation of the Role of Autophagy in Patients with Endometriosis of Different Stages during Secretory Phase and Proliferative Phase.

BACKGROUND: Autophagy exists widely in various physiological and pathological conditions. Lots of investigations have verified that the autophagic activity is always related to the occurrence and the development of cancer. Endometriosis (EMs) is a disease that endometrium-like tissues abnormally grow outside the uterus and also considered to possess the characters of tumor because of its malignant biological behavior. INTRODUCTION: Recently, several studies have already revealed that autophagy may play a potential role in proliferative-phase EMs. However, the function of autophagic activity in secretory-phase EMs is still unclear. METHODS: In our work, we explored autophagic activity between normal endometrium and EMs lesion endometrium during different menstrual phases and EMs stages. The clinical endometrium samples from 73 women were selected in this study, including 30 healthy individuals and 43 patients with EMs (endometrium samples include eutopic and its matched ectopic endometrium). All the participants were divided into two groups according to the menstrual cycle, namely proliferative-phase and secretive- phase group. Among the patients with EMs, 22 individuals in proliferative phase and the other 21 individuals in secretory phase were further classified into the groups of Stage I-II and Stage III-IV according to revised-American Fertility Society (r-AFS). Two autophagy-related proteins microtubuleassociated protein 1 light chain 3 beta-II (LC3B-II) and sequestosome protein (P62), which are believed to be the indicators of autophagy activity, were chosen in the study. Immunohistochemical (IHC) staining, Western blot assay and Real-Time quantitative Polymerase Chain Reaction (RTqPCR) were used to examine the expression of LC3B-II and P62 in protein and mRNA level accordingly. RESULT: It showed that the expression of LC3B-II both in protein and mRNA level decreased and that of P62 increased in secretory phase of the healthy group (P<0.05), but showed no significant difference in ectopic and its eutopic endometrium group during proliferative and secretory phase (P>0.05). In addition, the expression of LC3B-II in ectopic endometrium group was significantly lower than that of its eutopic endometrium group (P<0.05), and the expression of P62 was significantly higher accordingly (P<0.05). At the same time, both LC3B-II and P62 levels remained same between eutopic endometrium group and control group (P>0.05). Furthermore, compared to Stage I-II EMs group, the expression of LC3B-II was significantly lower (P<0.05) and P62 was significantly higher (P<0.05) in Stage III-IV EMs during secretory phase. CONCLUSION: Taken together, the periodicity-losing in EMs and the decreased autophagic activity in ectopic endometrium may exert a potential role in the pathogenesis of EMs. Down-regulated autophagy of ectopic endometrium in secretory phase may be related to the progression of EMs.

[Effects of DCC gene on cell growth and chemosensitivity of ovarian epithelial carcinoma cells].

OBJECTIVE: To study the effects of DCC gene transfection on cell-growth and chemosensitivity of ovarian epithelial carcinoma cell line HO8910. METHODS: Recombinant eukaryotic expression vector pcDNA3.1(+)-DCC containing DCC gene was introduced by lipofectamine transfection reagent into ovarian epithelial carcinoma cell line HO8910 which does not express DCC endogenously. The expression of DCC was detected by RT-PCR and immunocytochemistry. The cell proliferation and the viability rate after different concentrations of cisplatin and paclitaxel were given were assessed by methyl thiazolyl tetrazolium (MTT) assay. RESULTS: Exogenous DCC gene had been successfully transferred into HO8910 cells and obtained permanent expression. The growth speed of HO8910-DCC cells was significantly slower than other two groups. There was a significant difference between them (P < 0.01) except at the first day after being planted. There was no difference between the growth speed of HO8910 cells and that of HO8910-Neo cells (P > 0.05). The viability rate of HO8910-DCC cells was significantly lower than other two groups after (0.1 - 5.0) peak plasma concentration (PPC) of cisplatin and paclitaxel were given (P < 0.01). The viability rate of HO8910-DCC cells was lower than other two groups after 10.0 PPC concentration of cisplatin was given (P < 0.05), but there was no difference between them after 10.0 PPC concentration of paclitaxel was given (P > 0.05). The viability rate of HO8910 cells was similar to HO8910-Neo cells after different concentrations of cisplatin and paclitaxel were given (P > 0.05). CONCLUSION: The DCC gene expression not only inhibits cell growth but also enhances the chemosensitity of ovarian epithelial carcinoma cell line HO8910.

Silencing CCAT2 inhibited proliferation and invasion of epithelial ovarian carcinoma cells by regulating Wnt signaling pathway.

Long non-coding RNA CCAT2 (colon cancer-associated transcript 2) is dysregulated in varieties of human tumors. However, the role of CCAT2 in epithelial ovarian carcinoma (EOC) is not yet known clearly. The aim of this study is to investigate the effects of CCAT2 on proliferation and invasion of EOC cells and the potential mechanisms by which CCAT2 functions. In the present paper, we found that knockdown of CCAT2 impaired cell proliferation and invasion in vitro. Furthermore, we also studied the role of CCAT2 in the modulation of Wnt/ß-catenin signaling pathway. Our results showed that knockdown of CCAT2 inhibited the expression of ß-catenin and the activity of TCF/LEF (T-cell factor/lymphoid enhancer factor) acting as a key transcription factor of Wnt/ß-catenin signaling pathway. In addition, we found that silencing CCAT2 down-regulated the expression of c-MYC and MMP-7. But, that was reversed by the treatment with LiCl (lithium chloride) which could activate canonical Wnt/ß-catenin signaling pathway. Taken together, these results indicate that CCAT2 may promote ovarian cancer progression, at least partly, through Wnt/ß-catenin signaling pathway. Thus, CCAT2 might represent a novel therapeutic target for ovarian cancer.

[Growth inhibitory effects of lipofectamine-mediated DCC gene on ovarian epithelial carcinoma].

OBJECTIVE: To study the inhibitory effects of lipofectamine-mediated deleted colorectal carcinoma gene on ovarian epithelial carcinoma (ovarian cancer) cell line SKOV3. METHODS: We constructed a recombinant eukaryotic expression vector pcDNA3.1 (+)-DCC containing exogenous human DCC cDNA and vector with neomycin resistance gene, which were introduced by lipofectamine-mediated gene transfection into SKOV3 cell line that does not express DCC endogenously, thus forming SKOV3/DCC. Therefore, the experimental cells were classified into SKOV3/DCC, SKOV3/Neo and SKOV3. By using reverse transcriptase-polymerase chain reaction and immunocytochemistry, the expression of DCC mRNA and its protein were examined. RESULTS: Exogenous DCC had successfully been transferred into SKOV3 cells and obtained permanent expression. The growth speed of SKOV3/DCC was slower than the other two groups, there was significant difference between them (P < 0.01). SKOV3/DCC clones number was 38 +/- 8, while SKOV3 and SKOV3/Neo were 192 +/- 8 and 186 +/- 10, respectively, there was significant difference between them (P < 0.01). The percentage of G(1) phase cells increased to 78.0%, which that of S phase decreased to 5.3% by analyzing cell cycle, there was significant difference between them (20.0% and 3.2%, P < 0.01). The ultrastructural changes of the cells were observed under electron microscope, revealing growth retardation. CONCLUSION: DCC gene played an important role in generation and development of ovarian carcinomas.