[Clinical effect of superior oblique re-tucking for recurrent or residual compensatory head position].

Objective: To evaluate the effects of re-tucking the superior oblique muscle on recurrent or residual compensatory head position. Methods: A retrospective case series study was conducted. 12 recurrent or residual compensatory head position patients (12 eyes) with congenital superior oblique palsy who underwent superior oblique re-tucking in Tianjin Eye Hospital from March 2015 to February 2021 were included. All patients had a history of superior oblique tucking procedure and showed signs of superior oblique muscle palsy without inferior oblique muscle overaction. During surgery, the Guyton forced duction test is used to evaluate the relaxation of the superior oblique muscle tendon, which affects the re-tucking length of the muscle.Their head position, vertical deviation, eye movement, fovea-disa angle, and Bielschowsky head tilt test were assessed pre-and post-surgery. Statistical analysis was performed using ttest and paired samples Wilcoxon signed rank test. Results: Out of the 12 patients, 8 were male and 4 were female, aged between 2 and 9 years. The initial surgery was done at age 6, with a superior oblique recession length of (7.17±1.03) mm. Recurrent head tilt occurred in 11 patients after (3.82±0.98) months postoperatively, and 1 patient had residual head tilt, with a followup period of six months or more. Ocular motility examination revealed underaction of the superior oblique muscle, positive Bielschowsky's head tilt test, and Guyton forced duction tese indicating relaxation of the paralyzed superior oblique muscle tendon. Scar adhesion was observed at the stop of the superior oblique muscle, as well as the previous sutures. The scar and the sutures around the stop of the superior oblique muscle were released, the mean re-tucking amount was(7.83±1.59)mm. Follow-up at 12 to 18 months postoperatively showed disappearance of compensatory head position, significant improvement in superior oblique muscle lag, normal ocular motility, and no occurrence of Brown syndrome. The results of Bielschowsky head tilt were negative in 9 cases and still positive in 3 cases after superior oblique re-tucking. The primary vertical deviation was 2.5 (2.0, 5.3) prism diopter pre-operatively and 1 (0, 1) prism diopter post-operatively, respectively. The difference was statistically significant (U=6.00, P<0.001). The total amount of FDA in both eyes was (-22.04±5.47)° and (-15.27±6.08)° pre-and post-operatively, respectively. The difference was statistically significant (t=2.87, P=0.009). All 12 patients have normal eye movement after superior oblique re-tucking procedure. All patients had no compensatory head position at last follow-up. Conclusions: Superior oblique re-tucking is suitable for patients with relaxation of the superior oblique muscle tendon and extrocular rotation as the main sign. It can effectively and safely correct the recurrent or residual compensatory head position after re-tucking the superior oblique muscle.

[Application of indocyanine green iris angiography in the evaluation of anterior segment blood supply in different strabismus patients].

Objective: To evaluate the perfusion features of the anterior segment in patients with different types of strabismus. Methods: A cross-sectional study. Sixteen strabismus patients (16 eyes) who received the examination of iris indocyanine green angiography (ICGA) in Tianjin Eye Hospital from November 2016 to December 2021 were enrolled and divided into two groups according to whether they had a history of extraocular muscle injury/rectus muscle surgery. All patients underwent routine ophthalmic examinations. Angiographic images were obtained by the anterior segment camera, and indicators such as arm to iris circulation time, whole iris filling time, regression onset time, and complete regression time were recorded. The independent sample t test or Mann-Whitney U test was used to compare iris perfusion aspects of the two groups, and the Pearson/Spearman correlation tests was used to analyze the correlation of arm to iris circulation time and whole iris filling time with age and course of strabismus. Results: Among the 16 patients, there were 10 males and 6 females. The mean age was (49.2±13.2) years, and the course of strabismus ranged from 2 to 31 months. There were 7 patients in the group of without extraocualr muscle injury and 9 patients in the extraocular muscle injury/surgery group. There was no significant difference in age and course of strabismus between the two groups (both P>0.05). The arm to iris circulation time [M (Q1, Q3)] of the group without extraocular muscle injury and the group with extraocular muscle injury/surgery were 18 (18, 21) and 22 (20, 24) s, respectively. The average whole iris filling time was (13.86±1.95) and (12.22±3.60) s, respectively. There was no statistical significance between the two groups (both P>0.05). Correlation analysis showed that arm to iris circulation time was not correlated with age and course of strabismus (r=-0.033, -0.079; both P>0.05). And the whole iris filling time was not correlated with age and course of disease (r=0.057, -0.119; both P>0.05). The matrix scatter plots showed that in the group of extraocular muscle injury/surgery, there were three patients who were older than the average (49.2 years) and above the median of arm to iris circulation time (20 s) (two cases with the vertical muscle involved), meanwhile, there were three patients (all with the vertical muscle involved) whose course of strabismus was longer than 6 months and above the median of arm to iris circulation time, which were more than those in the group of without extraocular muscle injury (1 case, respectively). Conclusions: ICGA in patients with strabismus show that a history of injury to the extraocular muscle or surgical treatment beyond 2 months had no effect on iris perfusion. Age and course have no correlation with iris reperfusion. The vertical muscle involvement has more effects on the blood supply to the anterior segment.

[Graded vertical rectus tenotomy for small angle vertical deviation].

Objective: To observe the effectiveness of the graded vertical rectus tenotomy procedure for small-angle vertical deviation. Methods: Retrospective case series study. Twelve patients, including 8 males and 4 females, with an average age of (48±8) years were treated in Tianjin Eye Hospital from January 2017 to December 2019 for diplopia in primary gaze by strabismus surgery. The disease duration was (15±7) months. MRI/CT scan of the orbits and brain was performed to exclude the orbital and craniocerebral diseases. All patients underwent ocular movement examination, with the prism and alternate cover test to detect the deviation angle in primary gaze and the double Maddox test. Based on the results, the posterior segment of the nasal/temporal superior/inferior rectus muscle was operated. The changes of vertical and cyclotorsion deviation angle and the relationship between the vertical rectus graded tenotomy and corrected vertical deviation angle were observed at 1 day after surgery and the last follow-up. The vertical deviation angle was represented by M (Q1, Q3). Friedman test (Bonferroni correction) and linear fitting analysis were used for statistical analysis. Results: The follow-up time of all patients was (7±3) months. The vertical deviation angle in primary gaze before surgery [7.00 (5.25, 7.75) PD] was significantly different from that at 1 day after surgery [1.00 (1.00, 2.00) PD] and the last follow-up [1.50 (1.00, 2.00) PD] (P<0.001, P=0.003). There was no significant difference in postoperative cyclotorsion in all patients. Linear fitting results showed that 50% to 90% vertical rectus tenotomy corrected 5 to 8 PD vertical deviation (R2=0.72; P<0.001). Conclusion: The graded vertical rectus tenotomy procedure can effectively improve the small-angle vertical deviation in primary gaze.

[Clinical observation of anterior tucking of the superior oblique tendon for correction of excyclotropia in adults].

Objective: To investigate the clinical effect of anterior 1/3 superior oblique tendon tucking in acquired symptomatic excyclotropia adults. Methods: Retrospective case series. Seven patients (7 eyes) with acquired symptomatic excyclotropia who underwent an anterior 1/3 superior oblique tendon tuck procedure in Tianjin Eye Hospital from January 2019 to December 2019 were included. There were 5 male patients and 2 female patients, with an average age of (49±12) years old. All the patients had a history of a closed head injury. There was torsional diplopia in primary position, and eye movement examination showed paralysis of the superior oblique muscle in the affected eye, without obvious hyperactivity of the inferior oblique muscle. The anterior 1/3 tendon of the superior oblique muscle was tucked during surgery according to the relaxation of the superior oblique tendon in the forced duction test. The paired-sample nonparametric rank sum test was used to analyze the preoperative and postoperative (at 1 day after surgery and the last follow-up) excyclotropia angle. Results: The mean tuck amount was (7.7±1.8) mm (range, 6.0 to 10.0 mm). During operation, patients complained that excyclotropia improved markedly. The excyclotropia angle with the double Maddox rod test improved significantly from preoperative 10° (8°, 15°) to 2° (0°, 3°) at 1 day (Z=-2.379; P<0.05) and 2° (2°, 5°) at the last follow-up (Z=-2.375; P<0.05). The follow-up period was (112+38) days. All patients had no complaints of excyclotropia at the last follow-up. Conclusions: The anterior 1/3 superior oblique tendon tucking can effectively improve clinical symptoms in the primary position in adults with acquired excyclotropia and diplopia. The short-term postoperative results seem to be stable, without significant regression. (Chin J Ophthalmol, 2021, 57: 685-688).

[Clinical effects of partially de-epithelized local flaps in repairing tubercular chest wall defects].

Objective: To explore the clinical effects of partially de-epithelized local flaps in repairing tubercular chest wall defects. Methods: A retrospective observational study was conducted. From April 2010 to February 2021, twelve patients who met the inclusion criteria were admitted to the Department of Burns and Plastic Surgery of the Eighth Medical Center of PLA General Hospital, including 9 males and 3 females with age of (42±18) years. The sizes of tubercular chest wall defects of patients were ranged from 4 cm×3 cm×2 cm to 16 cm×8 cm×5 cm, which were all repaired with partial de-epithelized local flaps. The widths of flaps were equal to the widths of the defects, and the lengths of flaps were 2 cm longer than those of the defects. In one patient, the local flap was too large to close the donor site directly by suturing, so an autologous back free medium thickness skin graft was used for repair. In other patients, the collection areas of local flaps were small, and the donor areas of flaps were directly closed. The duration of operation, intraoperative bleeding, and postoperative drainage volume and indwelling time of drainage tube were observed and recorded. In two weeks after operation, the survival, color, and texture of flaps, the presence of subcutaneous hydrops and skin ulcer, and donor site healing including wound disruption, local infection, hematoma were observed. Chest X-ray, CT scan, or nuclear magnetic resonance imaging was performed in one month after operation to check whether new local hydrops and bone destruction occurred in the chest wall defects and the concomitant tuberculose focus of patients. All patients were followed up for more than 6 months to record whether the surgical incisions of the chest wall defects of the patients were complicated by hypertrophic scar, redness, swelling, and sinus. Results: In surgery, the patient had (104±18) min of operation duration, (119±53) mL of intraoperative bleeding, (134±49) mL of cumulative drainage of drainage tube, and (5.3±1.7) days of drainage tube indwelling time. In two weeks after operation, all the grafted local flaps survived, and the color and texture of flaps were similar to the surrounding normal skin. One patient had fluid leakage from the incision of chest wall defect area with the incision partially dehisced, which healed well after a phase Ⅱ operation; no wound infection, subcutaneous hydrops, or wound rupture occurred in other patients. The incisions of donor sites in all the patients healed well and no wound disruption, local infection, or hematoma occurred. One month after operation, no new bone destruction was observed in the operative region by chest imaging examination. Patients were followed up for 6 to 96 months, with one patient having wound swelling, ulceration, and sinus in the operative area of the chest wall defect in 12 months after surgery, which healed after phase Ⅱ operation; the incisions of chest wall defect wounds in other patients healed well and had no scar, redness and swelling, or sinus. Conclusions: Partially de-epithelized local flap could be used in repairing tubercular chest wall defect wounds, with the advantages of flexible flap design, minimal donor site injury, and good postoperative wound healing.

[Screening of biomarkers in exhaled breath of mice exposed to benzene].

Objective: To screen the biomarkers in the exhaled breath of mice exposed to benzene by using exhaled breath online analysis system. Methods: Thirty 8-week-old male C57BL/6 mice were randomly divided into six groups (0, 3, 32, 324, 648, and 1 296 mg/m3) and treated with benzene vapour for 28 days. At the end of the exposure, the peripheral blood cell counts and blood glutathione (GSH) were detected. The content of malondialdehyde (MDA) in HL60 cells treated by mice plasma was examined. Exhaled breath data from mice were collected by Secondary electrospray ionization source high resolution mass spectrometry (SESI-HRMS). Targeted analysis underlying benzene metabolites and oxidative stress metabolites was performed to screen the biomarkers in exhaled breath. Results: After benzene exposure, the number of peripheral blood cells was decreased in different degrees, particularly in the white blood cells (WBC) number. The WBC in 32 and 324 mg/m3 groups was declined by 27.76% and 52.87%, respectively compared to that in control group (P<0.05). Meanwhile, compared with the control group, the GSH content of peripheral blood cells from 324 mg/m3 group decreased by 13.16% (P<0.05). In addition, MDA content was increased by 18.11% in HL60 cells treated with plasma from 324 mg/m3 group mice (P<0.05). The phenol, hydroquinone/catechol, benzenetriol and trans, trans-Muconic acid (t,t-MA) in the exhaled gas of mice could be used as biomarkers for benzene exposure (R2>0.8, P<0.001). The peak intensity of five small molecular metabolites related to oxidative stress (ω-carboxylic fatty acid C5H10O3, ω-carboxylic fatty acid C6H12O3, glutamate, cysteine and MDA) increased with the increase of benzene concentration (P<0.05), which was negatively correlated with WBC decline (P<0.001), suggesting that these molecules mignt be used as biomarkers of benzene-induced toxicity. Conclusions: Phenol, hydroquinone/catechol, benzenetriol and trans, trans-Muconic acid (t,t-MA) in exhaled breath of mice could be used as biomarkers for benzene exposure; ω-carboxylic fatty acid C5H10O3, ω-carboxylic fatty acid C6H12O3, glutamate, cysteine and MDA might be used as markers of benzene-induced toxicity.

Niraparib maintenance therapy in patients with platinum-sensitive recurrent ovarian cancer using an individualized starting dose (NORA): a randomized, double-blind, placebo-controlled phase III trial☆.

BACKGROUND: This study evaluated maintenance treatment with niraparib, a potent inhibitor of poly(ADP-ribose) polymerase 1/2, in patients with platinum-sensitive recurrent ovarian cancer. PATIENTS AND METHODS: In this phase III, double-blind, placebo-controlled study conducted at 30 centers in China, adults with platinum-sensitive recurrent ovarian cancer who had responded to their most recent platinum-containing chemotherapy were randomized 2 : 1 to receive oral niraparib (300 mg/day) or matched placebo until disease progression or unacceptable toxicity (NCT03705156). Following a protocol amendment, patients with a bodyweight <77 kg or a platelet count <150 × 103/µl received 200 mg/day, and all other patients 300 mg/day, as an individualized starting dose (ISD). Randomization was carried out by an interactive web response system and stratified by BRCA mutation, time to recurrence following penultimate chemotherapy, and response to most recent chemotherapy. The primary endpoint was progression-free survival (PFS) assessed by blinded independent central review. RESULTS: Between 26 September 2017 and 2 February 2019, 265 patients were randomized to receive niraparib (n = 177) or placebo (n = 88); 249 patients received an ISD (300 mg, n = 14; 200 mg, n = 235) as per protocol. In the intention-to-treat population, median PFS was significantly longer for patients receiving niraparib versus placebo: 18.3 [95% confidence interval (CI), 10.9-not evaluable] versus 5.4 (95% CI, 3.7-5.7) months [hazard ratio (HR) = 0.32; 95% CI, 0.23-0.45; P < 0.0001], and a similar PFS benefit was observed in patients receiving an ISD, regardless of BRCA mutation status. Grade ≥3 treatment-emergent adverse events occurred in 50.8% and 19.3% of patients who received niraparib and placebo, respectively; the most common events were neutrophil count decreased (20.3% versus 8.0%) and anemia (14.7% versus 2.3%). CONCLUSIONS: Niraparib maintenance treatment reduced the risk of disease progression or death by 68% and prolonged PFS compared to placebo in patients with platinum-sensitive recurrent ovarian cancer. Individualized niraparib dosing is effective and safe and should be considered standard practice in this setting.

Influence of miR-155 on behaviors of depression mice through regulating Wnt/ß-catenin signaling pathway.

OBJECTIVE: To study the influence of micro ribonucleic acid (miR)-155 on depression-like behaviors of depression mice, and to explore the role of Wnt/b-catenin signaling pathway in behavioral regulation of depression mice. MATERIALS AND METHODS: The mouse model of depression was established via chronic unpredictable mild stress (CUMS). All mice were randomly divided into control group (n=12), model group (n=12), and fluoxetine group (n=12). The expression level of miR-155 in the hippocampus of mice in each group was detected via quantitative Polymerase Chain Reaction (qPCR). The changes in the behaviors of mice in each group were evaluated via behavioral experiments. The apoptosis level in the hippocampus of mice in each group was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Moreover, the content of inflammatory factors in the hippocampus of mice in each group was detected using the enzyme-linked immunosorbent assay (ELISA) kits. The expression levels of Wnt/b-catenin signaling pathway-related proteins in each group were detected via Western blotting. RESULTS: The expression level of miR-155 in the hippocampus was significantly higher in model group than that in control group (p<0.01). Meanwhile, the expression level of miR-155 was significantly lower in fluoxetine group than that in model group (p<0.01). There were no statistically significant differences in the crossing score and rearing score in the open field test among groups (p>0.05). Compared with those in control group, the immobility time in tail suspension test and forced swimming test were significantly increased (p<0.01), while the sucrose preference degree significantly declined (p<0.01) in model group. Fluoxetine could significantly reduce the immobility time in tail suspension test and forced swimming test (p<0.01) and increase the sucrose preference degree (p<0.01) in model group. The number of TUNEL-positive cells in the hippocampus of mice in model group was significantly larger than that in control group (p<0.01). Fluoxetine could effectively reduce the number of TUNEL-positive cells in the hippocampus (p<0.01). Compared with those in control group, the content of tumor necrosis factor-α (TNF-a), interleukin-1b (IL-1b), and IL-6 in the hippocampus was significantly increased (p<0.01), while the content of IL-10 was significantly decreased (p<0.01) in model group. Fluoxetine could effectively reduce the content of TNF-a, IL-1b, and IL-6 (p<0.01) and increase the content of IL-10 (p<0.01). Besides, in model group, the expression levels of dishevelled-1 (DVL-1) and b-catenin in hippocampus remarkably declined (p<0.01), while the expression levels of glycogen synthase kinase-3b (GSK-3b) and adenomatous polyposis coli (APC) were remarkably increased (p<0.01) compared with those in control group. Fluoxetine could effectively lower the expressions of GSK-3b and APC in the hippocampus (p<0.01) and increase the expressions of DVL-1 and b-catenin (p<0.01) in model group. CONCLUSIONS: MiR-155 is involved in regulating the depression-like behaviors of depression mice through promoting the release of inflammatory factors and the apoptosis of hippocampal neurons. Its mechanism may be related to the inhibition of the Wnt/b-catenin signaling pathway.

Effects of lncRNA BANCR on endometriosis through ERK/MAPK pathway.

OBJECTIVE: To investigate the regulatory role of long non-coding ribonucleic acid (lncRNA) BRAF-activated non-coding RNA (BANCR) in rats with endometriosis (EMs) and its mechanism of action. MATERIALS AND METHODS: A total of 30 healthy, unmated, female Sprague-Dawley (SD) rats were selected and divided into sham-operation group, model group and lncRNA BANCR intervention group, and a rat model of EMs was established by means of autotransplantation. The volume of eutopic endometrium in each group of rats was measured, and hematoxylin and eosin (HE) staining was applied to detect the impacts on the pathological morphology of ectopic endometrial tissues in each group. The levels of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and MMP-9 in the rat serum were determined by virtue of enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR) was performed to measure the messenger RNA (mRNA) levels of extracellular signal-regulated kinase (ERK) and mitogen-activated protein kinase (MAPK) in the uterine tissues in each group of rats, and Western blotting assay was adopted to detect the levels of phosphorylated ERK and MAPK proteins in the rat uterine tissues in each group. RESULTS: Compared with those in sham-operation group, the volume of eutopic endometrium in the rats was increased markedly, the pathological morphology was poorer, and the content of VEGF, MMP-2 and MMP-9 in the serum, the mRNA levels of ERK and MAPK in the uterine tissues, and the levels of phosphorylated ERK and MAPK proteins were elevated notably in model group. The rats in lncRNA BANCR intervention group had evidently decreased volume of eutopic endometrium, improved pathological morphology and significantly declined content of serum VEGF, MMP-2 and MMP-9, ERK and MAPK mRNA levels, and phosphorylated ERK and MAPK protein levels in the uterine tissues than those in model group. CONCLUSIONS: LncRNA BANCR inhibitor can repress the development of ectopic endometrial tissues by inhibiting the generation of angiogenic factors in the EMs focus, and its mechanism may be related to the inhibition on the ERK/MAPK signaling pathway.

MiR-15a-3p suppresses the growth and metastasis of ovarian cancer cell by targeting Twist1.

OBJECTIVE: To investigate the roles of miR-15a-3p in ovarian cancer cell growth and metastasis. PATIENTS AND METHODS: A key role of miR-15a-3p was identified via gene profiling and bioinformatics analysis. The impact of miR-15a-3p on ovarian cancer cell growth, migration and invasion was measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), wound-healing and transwell invasion assays. Bioinformatics and luciferase reporter assays were applied to identify that twist family BHLH transcription factor 1 (Twist1) was the target gene of miR-15a-3p. The miR-15a-3p level and the expression of Twist1 were detected using quantitative Real-time polymerase chain reaction (qRT-PCR) assay. The expressions of N-cadherin and E-cadherin were measured by immunofluorescence staining. Small interfering RNA targeting Twist1 and pCDNA3.1 containing Twist1 were applied to decrease and increase the expression of Twist1, respectively. RESULTS: miR-15a-3p was markedly down-regulated in ovarian cancer. Exogenous up-regulation of miR-15a-3p inhibited the growth, colony formation, migration and invasion of ovarian cancer cell in vitro. Furthermore, a xenograft model indicated that miR-15a-3p inhibited tumour growth and the metastatic potential of ovarian cancer cell in vivo. We found that Twist1 was the direct target of miR-15a-3p in ovarian cancer and that its expression was negatively correlated with the level of miR-15a-3p in ovarian cancer tissues. Up-regulation of miR-15a-3p rescued the inhibitory impact of miR-15a-3p on ovarian cancer cell growth, migration and invasion. Finally, down-regulation of Twist1 mimicked the suppressive effects of miR-15a-3p on ovarian cancer cell. CONCLUSIONS: We demonstrated that miR-15a-3p is down-regulated in ovarian cancer. Up-regulation of miR-15a-3p restrains the growth and metastasis of ovarian cancer cell by regulating Twist1.

Neuroprotective effects of α-lipoic acid on long-term experimental autoimmune encephalomyelitis.

OBJECTIVE: Experimental autoimmune encephalomyelitis (EAE) is an animal model commonly used in research on the acute phase of multiple sclerosis (MS), but studies on the pathology and pathogenesis of EAE with a long disease course are seldom conducted. Besides its antioxidant properties, the comprehensive mechanisms through which α-lipoic acid (LA) affects EAE remain obscure. We here conducted the study to explore the possible mechanisms. MATERIALS AND METHODS: In this study, the following methods were used for investigating the effects of LA on long-term EAE: hematoxylin-eosin staining (HE) and electron microscopic examinations of pathological changes; Western blotting of ß-amyloid precursor protein (ß-APP) and myelin basic protein (MBP); Enzyme-linked immunosorbent assay (ELISA) of tumor necrosis factor-α (TNF-α), transforming growth factor-ß (TGF-ß), superoxide dismutase (SOD), malondialdehyde (MDA) as well as flow cytometry of CD4+CD25+FoxP3+ regulatory T cells (Tregs). RESULTS: The results showed: (1) diverse pathological features of long-term relapsing-remitting EAE; (2) relatively increased MBP and reduced ß-APP expression in LA recipients 180 days after onset; (3) down-regulated TNF-α and up-regulated TGF-ß levels in LA recipients 7 days after onset; (4) lower MDA and higher SOD levels in LA recipients 180 days after onset; (5) increased Treg levels in LA recipients 7 days after onset. CONCLUSIONS: Aside from oxidative stress, LA possessed anti-inflammatory and immunomodulatory effects on EAE. LA might be a promising candidate for MS treatment.

Long non-coding RNA MALAT1 regulates ovarian cancer cell proliferation, migration and apoptosis through Wnt/ß-catenin signaling pathway.

OBJECTIVE: Long non-coding RNA (LncRNA) MALAT1 is an important regulatory molecule in many diseases, especially in ovarian cancer. We aimed at exploring the function of MALAT1 in ovarian cancer and at clarifying its mechanisms. PATIENTS AND METHODS: The expression level of MALAT1 in ovarian cancer tissues, para-carcinoma tissues and ovarian cancer cell lines were analyzed by Real-time polymerase chain reaction (RT-PCR). The cell proliferation rate was detected by CCK8 assay in SKOV3 and HO8910 cells. Transwell was used to detect the invasion and migration activities in SKOV3 and HO8910 cells. The cell cycle distribution and apoptosis rate were measured by flow cytometry analysis. The expression level of Dvl2, GSK-3ß, ß-catenin and cyclin D1 were detected by RT-PCR and Western blot. RESULTS: The relative expression level of MALAT1 was identified to be aberrantly up-regulated in ovarian cancer tissues and cell lines. The high expression level of MALAT1 was associated with poor prognosis in ovarian cancer patients. The down-regulation of MALAT1 inhibited cell proliferation, invasion and migration, arrested cell cycle progression in S phase and induced cell apoptosis in ovarian cancer cell lines. Meanwhile, the down-regulation of MALAT1 decreased the expression level of DVL2, ß-catenin and cyclin D1 and increased the expression level of GSK-3ß in SKOV3 and HO8910 cells. Moreover, the inhibitory effect of MALAT1 down-regulation in cell invasion and migration was reversed by SKL2001 activating Wnt/ß-catenin signal pathway and enhanced by XAV939 inhibiting Wnt/ß-catenin signal pathway. CONCLUSIONS: MALAT1 was overexpressed in ovarian cancer and associated to the poor prognosis. The down-regulation of MALAT1 inhibited cell proliferation, invasion and migration, arrested cell cycle progression in S phase and induced cell apoptosis by restraining the activation of Wnt/ß-catenin signaling pathway in ovarian cancer cells.

The diagnostic value of three detection techniques of ultrasound in the active stage of Crohn's disease.

OBJECTIVE: The widespread availability of Ultrasound machines and their relatively low cost and functionality make them an attractive tool to use during the treatment of ongoing Crohn's Disease (CD). This study aims at exploring the value of conventional, power Doppler and contrast-enhanced ultrasound during the active stages of CD. PATIENTS AND METHODS: 24 patients in the active stages of Crohn's disease were enrolled in the study. The full, medial and lateral intestinal wall thicknesses and the thickness ratio of medial to lateral intestinal wall of the segmental lesions were measured by conventional ultrasound. The diseased intestinal wall was also examined by power Doppler ultrasound to assign Limberg classification types: 3 cases were Limberg II, 9 cases were Limberg III and 12 cases were Limberg IV type. Importantly, the full and medial thicknesses of the intestinal walls with different Limberg types were compared, and statistically significant differences were found (p<0.05). Finally, images of the diseased segments were taken by contrast-enhanced ultrasound, and the contrast agent bolus arrival, the inflow and the peak enhancement times were calculated in order to be able to distinguish intestinal wall thickness differences according to different Limberg types. RESULTS: Cases with Limberg types III and IV mostly showed total intestinal enhancement, while Limberg II type cases showed mostly medial intestinal enhancement. When comparing the inflow and peak times of contrast-enhanced ultrasound of patients with different Limberg types, the differences found were statistically significant (p<0.05). CONCLUSIONS: This study confirms incrassation of the intestinal wall being the main ultrasonic appearance of active CD. Both power and contrast-enhanced ultrasound are effective tools in the management of active CD: while power ultrasound can be used to carry out Limberg typing, contrast-enhanced ultrasound can analyze and diagnose incrassation segments of the intestinal wall with different disease stages.

[Value of DNA content in comet tail in predicting changes in blood cell counts in workers exposed to benzene].

Objective: To investigate the value of DNA content in comet tail (TailDNA) in predicting the changes in peripheral blood cell counts in workers exposed to benzene. Methods: In 2011, cluster sampling was used to select 150 male workers exposed to benzene in a petrochemical factory. Cubital venous blood and urine samples were collected for routine blood rest, comet assay, and measurement of s-phenylmercapturic acid (SPMA) and urine creatinine. The population was divided into groups according to urinary SPMA or TailDNA, and routine blood test results within 3 years were collected to analyze the changes in blood cell counts. Results: The low-SPMA group had significantly higher white blood cell and neutrophilcounts in all years than the high-SPMA group (P<0.05) . The low-Tail DNA group had a significant increase in platelet count from 2012 to 2014 (P<0.05) , while the high-Tail DNA group had no significant change (P>0.05) . During the 4-year period, the high-TailDNA group had a significantly lower red blood cell count than the low-TailDNA group (P<0.05) . The high-TailDNA group showed a gradual reduction in white blood cell count over time (ß=-0.113, P<0.05) , and the low-TailDNA group showed no trend of the reduction in white blood cell count (P>0.05) . Conclusion: TailDNA can be used to predict the changes in blood cell counts in workers exposed to benzene.

[The effect of superior oblique tucking on the Bielschowsky head tilt test].

OBJECTIVE: To investigate the effect of superior oblique tucking on the Bielschowsky head tilt test. METHODS: A retrospective analysis of 22 patients with congenital unilateral superior oblique palsy, who underwent tucking of the superior oblique tendon in Tianjin Eye Hospital, depending on vertical deviation in the primary position, ocular motility, objective cyclotorsion and abnormal head posture. Bielschowsky head tilt test was evaluated quantitatively by the prism cover test. Α positive Bielschowsky head tilt test was defined as one in which the vertical deviation when the head tilt to the side of the paresis was at least 5. 0(△) greater than that on tilt to the uninvolved side. The follow-up period ranged from 3 to 12 months (mean 4.6 months). Kolmogorov-Smirnov (K-S), Rank sum test and Spearman correlation analysis statistical methods were used in this study. RESULTS: All patients had vertical deviation and significant abnormal head posture before operation. After the procedure of superior oblique tucking,vertical deviation and symptom of unacceptable abnormal head posture were ameliorated or disappeared. OBJECTIVE torsion was 15.62°±7.36° before the surgery, 9.91°±10.09°1d after the surgery and 11.25°±9.17°at the last follow-up visit, respectively (P<0.05). We found that there were positive correlations between the objective cyclotorsion and the vertical deviation difference value between the paralyzed side and uninvolved side at 1 d after the surgery and last follow-up visit(P<0.05). The vertical deviation of the paresised eye when the head tilted to the side of paresis side and uninvolved side difference were 5.00(△)-17.00(△), average 8.68(△)±3.23(△), 1.00(△)-8.00(△),average 3.36(△)±2.01(△) and 0.00(△)-14.00(△),average 3.77(△)±3.01(△), preoperatively, 1d after the surgery and at the last follow-up visit respectively. Using Rank sum test, the pre-and postoperative difference was statistically significant (Z=-4.64, P<0.01). A positive Bielschowsky head tilt test was found in all cases preoperatively. The results of Bielschowsky tilt test was still positive in 5 cases and negative in 17 cases (77.3%). CONCLUSIONS: In most cases with unilateral superior oblique paresis, the results of Bielschowsky tilt test became negative after superior oblique tucking. The long-standing results is worthy of observation. (Chin J Ophthalmol, 2016, 52: 589-595).

SP6616 as a new Kv2.1 channel inhibitor efficiently promotes ß-cell survival involving both PKC/Erk1/2 and CaM/PI3K/Akt signaling pathways.

Kv2.1 as a voltage-gated potassium (Kv) channel subunit has a pivotal role in the regulation of glucose-stimulated insulin secretion (GSIS) and pancreatic ß-cell apoptosis, and is believed to be a promising target for anti-diabetic drug discovery, although the mechanism underlying the Kv2.1-mediated ß-cell apoptosis is obscure. Here, the small molecular compound, ethyl 5-(3-ethoxy-4-methoxyphenyl)-2-(4-hydroxy-3-methoxybenzylidene)-7-methyl-3-oxo-2,3-dihydro-5H-[1,3]thiazolo[3,2-a]pyrimidine-6-carboxylate (SP6616) was discovered to be a new Kv2.1 inhibitor. It was effective in both promoting GSIS and protecting ß cells from apoptosis. Evaluation of SP6616 on either high-fat diet combined with streptozocin-induced type 2 diabetic mice or db/db mice further verified its efficacy in the amelioration of ß-cell dysfunction and glucose homeostasis. SP6616 treatment efficiently increased serum insulin level, restored ß-cell mass, decreased fasting blood glucose and glycated hemoglobin levels, and improved oral glucose tolerance. Mechanism study indicated that the promotion of SP6616 on ß-cell survival was tightly linked to its regulation against both protein kinases C (PKC)/extracellular-regulated protein kinases 1/2 (Erk1/2) and calmodulin(CaM)/phosphatidylinositol 3-kinase(PI3K)/serine/threonine-specific protein kinase (Akt) signaling pathways. To our knowledge, this may be the first report on the underlying pathway responsible for the Kv2.1-mediated ß-cell protection. In addition, our study has also highlighted the potential of SP6616 in the treatment of type 2 diabetes.

Association of the genetic polymorphisms of NFKB1 with susceptibility to ovarian cancer.

Nuclear factor-κB (NF-κB), a transcription factor that is activated by various stimuli, is associated with the pathogenesis of several cancers. One functional polymorphism, -94 insertion/deletion ATTG (rs28362491), in the human NFKB1 gene (one member of the NF-κB gene family) is associated with increased risk of various cancers. However, only one study has reported that rs28362491 is significantly associated with ovarian cancer. The aim of this study was to analyze the association between single nucleotide polymorphisms (SNPs) and haplotypes in the NFKB1 gene and the risk of ovarian cancer in a Chinese population. We examined the potential association between ovarian cancer and 15 SNPs (rs28362491, rs3774932, rs1598856, rs230531, rs230530, rs230528, rs230521, rs230498, rs230539, rs1005819, rs3774956, rs4648055, rs4648068, rs3774964, rs3774968) of the NFKB1 gene using the MassARRAY system. Participants included 411 patients with ovarian cancer and 438 healthy controls. The results showed that the allelic or genotypic frequencies of three polymorphisms, including rs28362491 (promoter region), rs230521 (intron 4), and rs4648068 (intron 12), in the patients with ovarian cancer, were significantly different from those in the healthy controls. Strong linkage disequilibrium was observed in four blocks (D' > 0.9). Significantly more A-C (block 2: rs230528-rs230521) haplotypes (P = 0.0003 after Bonferroni's corrections) and G-A-A (block 4: rs4648068-rs3774964-rs3774968) haplotypes (P = 0.021) were found in the patients with ovarian cancer. These findings point to a role of the NFKB1 polymorphism in patients with ovarian cancer among a Chinese Han population, and may be informative for future genetic or biological studies on ovarian cancer.

Association between dietary intake of folate and MTHFR and MTR genotype with risk of breast cancer.

We investigated the association between dietary intake of folate, vitamin B6, and the 5,10-methylenetetrahydrofolate reductase (MTHFR) genotype with breast cancer. A matched case-control study was conducted, and 413 patients with newly diagnosed and histologically confirmed breast cancer and 436 controls were recruited. Folate intake, vitamin B6, and vitamin B12 levels were calculated, and the MTHFR C677T and A1298C and MTR A2756G polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism. Breast cancer cases were generally older, older at first live birth, and younger at menarche, had a higher body mass index, were smokers, had higher energy intake, and more first-degree relatives with breast cancer as well as more live births compared to controls. With respect to energy intake, we found that higher energy intake were more likely to increase the risk of breast cancer. The MTHFR 667TT genotype was associated with a moderately increased risk of breast cancer when compared with the CC genotype, and a significant odds ratio (OR; 95% confidence interval, CI) was found (OR = 1.70, 95%CI = 1.06-2.73). Individuals carrying T allele were associated with higher risk of breast cancer when compared with C allele (OR = 1.34, 95%CI = 1.06-1.70). We did not find a significant effect of the MTHFR A1298C and MTR A2756G on the risk of breast cancer. We did not find any association between folate intake and MTHFR C677T polymorphisms. In conclusion, we found that the MTHFR C667T polymorphism is associated with the risk of breast cancer, indicating that this genotype plays a role in breast cancer development.

Upregulation of miR-27a contributes to the malignant transformation of human bronchial epithelial cells induced by SV40 small T antigen.

The introduction of the Simian virus 40 (SV40) early region, the telomerase catalytic subunit (hTERT) and an oncogenic allele of H-Ras directly transforms primary human cells. SV40 small T antigen (ST), which forms a complex with protein phosphatase 2A (PP2A) and inhibits PP2A activity, is believed to have a critical role in the malignant transformation of human cells. Recent evidence has shown that aberrant microRNA (miRNA) expression patterns are correlated with cancer development. Here, we identified miR-27a as a differentially expressed miRNA in SV40 ST-expressing cells. miR-27a is upregulated in SV40 ST-transformed human bronchial epithelial cells (HBERST). Suppression of miR-27a expression in HBERST cells or lung cancer cell lines (NCI-H226 and SK-MES-1) that exhibited high levels of miR-27a expression lead to cell growth arrested in the G(0)-G(1) phase. In addition, suppression of miR-27a in HBERST cells attenuated the capacity of such cells to grow in an anchorage-independent manner. We also found that suppression of the PP2A B56γ expression resulted in upregulation of miR-27a similar to that achieved by the introduction of ST, indicating that dysregulation of miR-27a expression in ST-expressing cells was mediated by the ST-PP2A interaction. Moreover, we discovered that Fbxw7 gene encoding F-box/WD repeat-containing protein 7 was a potential miR-27a target validated by dual-luciferase reporter system analysis. The inverse correlation between miR-27a expression levels and Fbxw7 protein expression was further confirmed in both cell models and human tumor samples. Fbxw7 regulates cell-cycle progression through the ubiquitin-dependent proteolysis of a set of substrates, including c-Myc, c-Jun, cyclin E1 and Notch 1. Thus, promotion of cell growth arising from the suppression of Fbxw7 by miR-27a overexpression might be responsible for the viral oncoprotein ST-induced malignant transformation. These observations demonstrate that miR-27a functions as an oncogene in human tumorigenesis.

α4 is highly expressed in carcinogen-transformed human cells and primary human cancers.

A regulator of the protein phosphatase 2A (PP2A), α4, has been implicated in a variety of functions that regulate many cellular processes. To explore the role of α4 in human cell transformation and tumorigenesis, we show that α4 is highly expressed in human cells transformed by chemical carcinogens including benzo(a)pyrene, aflatoxin B(1), N-methyl-N'-nitro-N-nitrosoguanidine, nickel sulfate and in several hepatic and lung cancer cell lines. In addition, overexpression of α4 was detected in 87.5% (74/80) of primary hepatocellular carcinomas, 84.0% (21/25) of primary lung cancers and 81.8% (9/11) of primary breast cancers, indicating that α4 is ubiquitously highly expressed in human cancer. Functional studies revealed that elevated α4 expression results in an increase in cell proliferation, promotion of cell survival and decreased PP2A-attributable activity. Importantly, ectopic expression of α4 permits non-transformed human embryonic kidney cells (HEKTER) and L02R cells to form tumors in immunodeficient mice. Furthermore, we show that the highly expressed α4 in transformed cells or human tumors is not regulated by DNA hypomethylation. A microRNA, miR-34b, that suppresses the expression of α4 through specific binding to the 3'-untranslated region of α4 is downregulated in transformed or human lung tumors. Taken together, these observations identify that α4 possesses an oncogenic function. Reduction of PP2A activity due to an enhanced α4-PP2A interaction contributes directly to chemical carcinogen-induced tumorigenesis.