[Effect of miR1290 on the growth of endometrial cancer and the related mechanism].

Objective: To explore the expression of miR1290 in endometrial cancer tissues and its relationship with the pathological grade, and to find out the effect of miR1290 on biological characteristics of endometrial cancer cells and its mechanism. Methods: A total of 38 cases of endometrioid adenocarcinoma tissues, 10 cases of adjacent tissues and 23 cases of normal endometrial tissues were collected in Provincial Hospital Affiliated to Shandong University from May 2020 to October 2020. The expression of miR1290 was detected by reverse transcription polymerase chain reaction (RT-PCR). The expressions of miR1290 in endometrial cancer cells including KLE and Ishikawa were knocked down by lentiviral transfection. Cell counting kit 8 (CCK-8) test and colony formation test were used to detect cell proliferation ability, wound healing and Transwell test were used to detect cell invasion and migration ability, western blot was used to detect the expressions of epithelial-mesenchymal transition (EMT), phospholipids acylinositide 3-kinase (PI3K)/Akt and Wnt/ß-catenin pathway related proteins. Results: The relative expressions of miR1290 in endometrial cancer tissues were 5.40±3.20, which was 1.55 times of normal endometrial tissues (P<0.01) and 1.75 times of adjacent tissues (P<0.01). The relative expressions of miR1290 in 17 cases of endometrial tissues at proliferative stage and 6 cases of endometrial tissues at secretory stage were 3.00±1.08 and 4.97±0.58, respectively, and the difference was statistically significant (P<0.01). In KLE cells and Ishikawa cells, the expression of miR1290 in miR1290 knockdown (Sh-miR1290) group was decreased when compared with the negative control (Sh-NC) group. The absorbance value of Sh-miR1290 group detected by the CCK-8 method and the colony formation rate detected by the colony formation experiment were both increased, the number of cells penetrating the basement membrane in the Transwell experiment and the wound healing rate in the scratch experiment were decreased (P<0.05). In KLE cells, knockdown of miR1290 reduced the expressions of EMT-related proteins including N-cadherin, Vimentin, Snail and Slug(P<0.05), and the expressions of PI3K and P-Akt/Akt (P<0.05), while there was no significant change in the expressions of Wnt and ß-catenin (P>0.05). In Ishikawa cells, knockdown of miR1290 decreased the expressions of EMT-related proteins including N-cadherin, Snail and Slug, and the expressions of Wnt and ß-catenin, increased the expression of E-cadherin (P<0.05), while there was no significant change in the expressions of PI3K and P-Akt/Akt (P>0.05). Conclusions: The expressions of miR1290 in endometrial cancer tissues are higher than that in the adjacent tissues and normal endometrial tissues. Knockdown of miR1290 expression can promote the proliferation of endometrial cancer cells, but inhibit cell invasion, migration and EMT ability through the PI3K/Akt and Wnt/ß-catenin pathways.

Application of Eye Tracker in Lie Detection.

ABSTRACT: Objective To investigate the application value of eye tracking in lie detection. Methods The 40 subjects were randomly divided into two groups. The pupil diameter, fixation duration, points of fixation and blink frequency of the subjects in the experimental group in observing target stimulation and non-target stimulation were recorded with eye tracker after they accomplished the mock crime. The eye movement parameters of subjects in the control group were directly collected. The differences in eye movement parameters of the experimental group and the control group in observing target stimulation and non-target stimulation were analyzed by t-test. Pearson coefficient analysis of correlation between eye movement parameters that had differences was conducted. The effectiveness of eye movement parameters to distinguish between the experimental group and the control group was calculated by the receiver operator characteristic （ROC） curve. Results Participants from the experimental group had shorter average pupil diameter, longer average fixation duration and fewer fixation points （P<0.05）, but the differences in blink frequency had no statistical significance. The differences in the above indicators of the control group in observing target stimulation and non-target stimulation had no statistical significance. The average fixation duration showed a negative correlation with fixation points （r=-0.255, P<0.05）; the average fixation duration showed a negative correlation with average pupil diameter （r=-0.218, P<0.05）; the fixation points showed a positive correlation with average pupil diameter （r=0.09, P<0.05）. The area under the curve of average pupil diameter, average fixation duration and fixation points was 0.603, 0.621 and 0.580, respectively. Conclusion The average pupil diameter, average fixation duration and fixation points obtained by the eye tracker under laboratory conditions can be used to detect lies.

Bioactive components in the fruits of Ziziphus jujuba Mill. against the inflammatory irritant action of Euphorbia plants.

Chinese jujube (also known as Chinese date) is the fruit of Ziziphus jujuba Mill. (Rhamnaceae). As a famous folk medicine, it is used as antidote in traditional Chinese formula, Shi Zao Decoction, to relieve the drastic inflammatory irritant nature of Euphorbia species. The irritant activities may cause serious adverse effects in clinical practices. This study aimed to investigate the active components of Z. jujuba through the inhibitory effects on the inflammatory cells activated by Euphorbia kansui and prostratin, a phorbol ester isolated from Euphorbia fischeriana. Peritoneal macrophage of rat and splenic lymphocyte (splenocyte) of mouse were selected to evaluate these actions in vitro. Nitric oxide (NO) release of macrophage and the proliferation of splenocyte were examined through Griess method and MTT assay. TNF-α, as an important pro-inflammatory cytokines, was detected with enzyme-linked immunosorbent assay (ELISA) method. Six fractions extracted from Z. jujuba were evaluated and fraction F (triterpene acids fraction) was demonstrated to be the most active part, and then, 21 compounds isolated from Z. jujuba were tested at the concentrations range from 1 µg/ml to 100 µg/ml. The results show that 7 compounds of them are likely to be active compounds concerning to their pronounced inhibitory action on the activated inflammatory cells. These effects might be helpful to attenuate the irritant action of Euphorbiaceae plants and protect the gastrointestinal tissue from potent inflammatory injury, which should be beneficial to some diseases, like inflammatory bowel disease.

Femtosecond laser-drilling-induced HgCdTe photodiodes.

Femtosecond-laser drilling may induce holes in HgCdTe with morphology similar to that induced by ion-milling in loophole technique. So-formed hole structures are proven to be pn junction diodes by the laser beam induced current characterization as well as the conductivity measurement. Transmission and photoluminescence spectral measurements on a n-type dominated hole-array structure give rise to different results from those of an ion-milled sample.

Effects of crocetin on the matrix metalloproteinases in cardiac hypertrophy induced by norepinephrine in rats.

The effects of crocetin on the cardiac hypertrophy induced by long-term treatment with norepinephrine (NE) in rats have been investigated. The activities of matrix metalloproteinases (MMP-2 and MMP-9) have been assayed by gelatin SDS-PAGE zymography. The expressions of MMP-2 and MMP-9 were detected by RT-PCR. ATPase activity and hydroxyproline contents were measured with a commercial kit. The results show that crocetin blocked the development of left ventricular hypertrophy induced by NE, decreased the level of collagen in myocardium, enhanced both the Na+-K+ ATPase activity in cardiac tissue and the Ca2+-Mg2+ ATPase activity in mitochondria and inhibited significantly the activity of MMP-2 and the expressions of MMP-2 and MMP-9. These results suggest that crocetin may prevent cardiac hypertrophy induced by NE in rats.

The effect of aging on distraction osteogenesis in the rat.

The effect of age on bone formation in the limb lengthening model of distraction osteogenesis (DO) was investigated in two studies using Sprague-Dawley (SD) rats from two colonies at various ages (CAMM: 9 vs 24 months, Harlan: 4 vs 24 months). External fixators were placed on the right tibiae of 30 male SD rats (20 CAMM, 10 Harlan) and mid-diaphyseal osteotomies were performed. Distraction was performed at 0.2 mm bid for 20 days (CAMM) or 14 days (Harlan). The experimental (DO) and control (contra-lateral) tibiae were removed for high-resolution radiography and decalcified histology. Videomicroscopy was used to quantitate radiodensity, histology (matrix type) and relative areas of cell proliferation, which was identified by proliferating cell nuclear antigen (PCNA) immunochemistry. Both studies demonstrated an age-related decrease in the percent mineralized bone (radiodensity) in the distraction gap (CAMM 9 vs 24 months: 68% vs 51%, P < 0.003; Harlan 4 vs 24 months: 95% vs 36%, P < 0.001) and no significant colony or distraction time-specific difference was seen between the two colonies of 24-month-old rats. Histology was performed on the Harlan rats. The DO gaps in the 24-month-old rats demonstrated less endosteal new bone compared to the 4-month-old rats (P < 0.01), but equivalent periosteal new bone. In 4-month-old rats, PCNA-immunostained cells were organized along the primary matrix front (where the first deposition of osteoid occurs) extending across both periosteal and endosteal surfaces. In 24-month-old rats, PCNA+ cells were organized in zones along the periosteal new bone fronts only and irregularly scattered throughout the endosteal gap within a fibrovascular non-ossifying matrix. These results indicate that 24-month-old rats have a relative deficit in endosteal bone formation which may not be related to cell proliferation but rather to cell organization. This model reflects the clinical situation where radiographic findings in older patients demonstrate significant delays in mineralization during DO. We believe this model of DO in aged rats presents unique in vivo opportunities to test hypotheses concerning (1) the effects of aging on bone repair, (2) the effects of pharmacological agents on bone repair in a geriatric setting, and (3) to study the mechanisms underlying DO.

Development of tensile strength during distraction osteogenesis in a rat model.

These studies were designed to determine the reliability of in vitro tensile testing to measure the temporal development of regenerate bone strength in rats during limb lengthening (distraction osteogenesis, DO). External fixators were placed on the right tibiae of 36 virus-free, 400-450 g male Sprague Dawley rats, and osteotomies (n = 33) were performed. Distraction was initiated the following morning (0 day latency) at 0.4 mm/day and continued to day 20. The 8 mm gap was allowed to consolidate for up to 50 days (day 70 postop). Contralateral unoperated and operated (fixator only) controls were included. On days 20, 30, 50 and 70 postop, the rats were anesthetized, and their tibiae were radiographed prior to undergoing sacrifice for histological or tensile analysis. On day 70, an additional group was tested by three-point bending. Radiodensity measurements demonstrated progressive mineralization of the DO gap, and histology confirmed typical intramembranous ossification of collagen bundles oriented parallel to the distraction force. Tensile stiffness increased significantly between days 20 and 30 postop, this increase correlated with initial radiographic and histologic bridging of the DO gap. Energy to failure and ultimate tensile strength increased progressively to day 70. At day 70, the force to failure for three-point bending was 65% of control tibiae. In conclusion, in vitro tensile testing provides a reliable method to test the development of structural integrity during the early stages of DO. Therefore, the biomechanical effects of postulated modulators of bone repair can be measured during early stages (bone formation, bridging, early consolidation) of DO in a rat model.

[Gene chimeric fusion and expression of nucleocapsid NS3 regions and NS4 regions of hepatitis C virus genome].

Genes encoding HCV core and NS4 antigen epitopes and C33c antigen were cloned from HCV genome by PCR, respectively. Two fused genes were constructed. One contained these three genes, another contained genes encoding C33c antigen and NS4 antigen epitopes. These fused genes were cloned into expression plasmid pET-24(a)+ and pET-22(b)+ under T7 promoter and transformed into E. coli BL21 (DE3) respectively. SDS-PAGE analysis revealed that these fused antigens CCN, CN were highly expressed after the induction by 1 mmol/L IPTG. These Expression products were detected by western blotting with anti-HCV serum.

Ribosomal RNA supplementation highly reinforced cell-free translation activity of wheat germ.

We have constructed an inexpensive, highly efficient eukaryotic cell-free translation system. Wheat germ rRNA (WG rRNA) was prepared by phenol/chloroform (P/C) extraction, a simple and quick method, from wheat germ, an inexpensive and commercially available by-product of flour production. Addition of a small amount of WG rRNA into a wheat germ cell-free translation system increased the protein productivity of the system 6- to 8-fold. Isolated 18S or 28S rRNA alone enhanced the protein production only 2-fold or 3.9-fold, respectively, at maximum. On the other hand, their equimolar mixture enhanced the production as much as the whole WG rRNA, indicating 18S and 28S rRNA synergistically functioned to enhance protein synthesis. Addition of WG rRNA slightly improved the stability of mRNA in the cell-free translation system, which explained only partly the enhancement of protein production. Addition of WGE or ribosome containing approximately the same amount of rRNA in the form of protein-rRNA complex as WG rRNA added to the system did not increase the protein production in the translation system. When ribosome in the cell-free translation system was replaced with WG rRNA, the system did not exhibit any detectable translation activity, indicating that the translation activity of WG rRNA is negligible in comparison with that of ribosome. These results indicated that WG rRNA affected some mechanisms regulating the translation rate in wheat germ cell-free system, resulting in increased protein production.

Sustained proliferation accompanies distraction osteogenesis in the rat.

These studies were conducted to compare the local cellular proliferation patterns in the rat tibia during distraction osteogenesis with those during nondistracted fracture healing. Bone specimens from distraction osteogenesis and nondistracted fracture groups were analyzed 2, 10, and 20 days after surgery. Proliferation was determined by metabolic labeling with [3H]thymidine and by immunocytochemistry with an antibody to proliferating cell nuclear antigen. Videomicroscopy was used to count the cells staining positively within specified regions. The number of cells incorporating [3H]thymidine was positively correlated (r2 = 0.78) with the number of proliferating cell nuclear antigen positive cells on alternating serial slides. At day 2, the latter cells were largely confined to the bone marrow and periosteum in both groups, and the cell numbers per mm2 were also equivalent. At days 10 and 20, the proliferating cell nuclear antigen positive cells predominated in both the proximal and distal primary matrix front zones in the distraction osteogenesis group. In contrast, the proliferating cell nuclear antigen positive cells in the nondistracted fracture group were scattered throughout the healing area. Significantly more of these cells were in the primary matrix front zones than in any location within the nondistracted fracture-healing area. The number of these cells in the bone marrow adjacent to the surgical area declined from day 2 to day 10 in both groups. The results suggest that (a) proliferating cell nuclear antigen immunostaining is a reliable indicator of cycling cells; (b) by day 10, distraction osteogenesis is characterized by a zone-specific pattern of proliferating cells; and (c) distraction osteogenesis prolongs the stimulation of proliferation within the gap after fracture.

Rat model of distraction osteogenesis.

Prior studies of distraction osteogenesis in dog and rabbit models have shown predominantly intramembranous bone formation. Other models of fracture healing normally display mixtures of both endochondral and intramembranous bone formation. We have established a rat model of tibial lengthening that reliably reproduces the pattern of zonal osteogenesis previously observed in dog and rabbit models. A distraction rate of 0.25 mm twice a day with a 0-day latency period produced intramembranous bone with zones of progressive mineralization from collagen. With this protocol, rats bridged the distraction gap with a 25% increase in the tibial bone length. After 20 days of distraction and 50 days of consolidation, the three-point bending stiffness, as a percentage of the contralateral control, reached a level equivalent to that measured in the canine model for a 15% lengthening (28-day distraction and 84-day consolidation). Radiodensitometric analysis of the regenerate bones measured 97% of the unaffected contralateral tibial densities, and mineral analyses demonstrated that calcium and phosphorus levels in the regenerate bone reached 78% of contralateral tibial levels by day 70. We concluded that a rat model of distraction osteogenesis will be useful for a wide range of studies involving rapid intramembranous bone formation.

The impact of total enteral nutrition on distraction osteogenesis in a rat model.

Limb lengthening by gradual mechanical distraction, termed distraction osteogenesis (DO), results in new bone formation. We have developed a rat tibial model for DO and have proceeded to study the effects of nutrition on this process. We have combined the intragastric diet delivery system of total enteral nutrition (TEN) with DO in the rat model. The first study was designed to address the weight loss associated with DO in dogs and patients. Rats in the chow + DO group lost 10% body weight over the 20-day distraction period but gradually gained weight back to the preoperative level by the end of the 5th week of the bone consolidation period. In contrast, in the TEN + DO group, a weight gain was recorded during the 20-day distraction phase. A second study was conducted to determine the effects of TEN on the rate and histology of regenerate bone formation. The weight changes replicated those seen in the first study. Standardized radiographs, taken on day 20, revealed increases (p < 0.003) in regenerate bone formation in the TEN group when compared with the chow group. Increased numbers of osteoclasts in the TEN group may indicate an accelerated entry into the remodeling phase of consolidation. Serum IGF-I values, taken at day 20, did not differ between the groups. These results demonstrate that the nutritional support dramatically increased the mineralized bone formed over the 20-day distraction period.

Cytokeratin expression in non-neoplastic oesophageal epithelium and squamous cell carcinoma of the oesophagus.

The expression of cytokeratins (CK) 19, 8, 18, 13, 10 and 7 was examined in 35 cases of squamous cell carcinomas of the oesophagus (10 well-differentiated, 13 moderately-differentiated, and 12 poorly-differentiated) and the adjacent mucosa by means of a panel of monoclonal antibodies on frozen sections. The study was undertaken to assess the pattern of expression of these keratins in oesophageal tumours and its relation to the degree of differentiation. The normal oesophageal epithelia expressed CK19 in 86%, CK18 in 17% and CK13 in 14% of cases. CK8, CK10 and CK7 immunoreactivity was not observed. The tumours expressed CK19 in 86%, CK8 in 46%, CK18 in 97%, CK13 in 83%, CK10 in 34% and CK7 in 29% of cases. Thus, the so-called simple epithelial markers CK18 and CK19 occurred in the majority of oesophageal squamous cell carcinomas. CK13 (the so-called non-keratinizing squamous epithelial marker) was only infrequently demonstrated in the non-neoplastic oesophageal mucosa, and its expression was more frequent in carcinomas. CK10 was not demonstrated in non-neoplastic mucosa, but was mostly associated with well-differentiated carcinomas. We therefore conclude that the pattern of expression of cytokeratins in oesophageal carcinomas is different from that in normal oesophageal epithelia and varies with differentiation.