Genome-Wide Identification of bZIP Transcription Factors in Faba Bean Based on Transcriptome Analysis and Investigation of Their Function in Drought Response.

Faba bean is an important cool-season edible legume crop that is constantly threatened by abiotic stresses such as drought. The basic leucine zipper (bZIP) gene family is one of the most abundant and diverse families of transcription factors in plants. It regulates plant growth and development and plays an important role in the response to biotic and abiotic stresses. In this study, we identified 18 members of the faba bean bZIP transcription factor family at the genome-wide level based on previous faba bean drought stress transcriptome sequencing data. A phylogenetic tree was constructed to group the 18 VfbZIP proteins into eight clades. Analysis of cis-acting elements in the promoter region suggested that these 18 VfbZIPs may be involved in regulating abiotic stress responses such as drought. Transcriptome data showed high expression of seven genes (VfbZIP1, VfbZIP2, VfbZIP5, VfbZIP7, VfbZIP15, VfbZIP17, and VfbZIP18) in the drought-tolerant cultivar under drought stress, in which VfbZIP1, VfbZIP2, and VfbZIP5 were consistently expressed as detected by quantitative real-time polymerase chain reaction (qRT-PCR) compared to the transcriptome data. Ectopic overexpression of the three VfbZIPs in tobacco, based on the potato Virus X (PVX) vector, revealed that VfbZIP5 enhanced the drought tolerance. Overexpressed VfbZIP5 in plants showed lower levels of proline (PRO), malondialdehyde (MDA), and peroxidase (POD) compared to those overexpressing an empty vector under 10 days of drought stress. Protein-protein interaction (PPI) analysis showed that VfbZIP5 interacted with seven proteins in faba bean, including VfbZIP7 and VfbZIP10. The results depict the importance of VfbZIPs in response to drought stress, and they would be useful for the improvement of drought tolerance.

Perspectives of herbs and their natural compounds, and herb formulas on treating diverse diseases through regulating complicated JAK/STAT signaling.

JAK/STAT signaling pathways are closely associated with multiple biological processes involved in cell proliferation, apoptosis, inflammation, differentiation, immune response, and epigenetics. Abnormal activation of the STAT pathway can contribute to disease progressions under various conditions. Moreover, tofacitinib and baricitinib as the JAK/STAT inhibitors have been recently approved by the FDA for rheumatology disease treatment. Therefore, influences on the STAT signaling pathway have potential and perspective approaches for diverse diseases. Chinese herbs in traditional Chinese medicine (TCM), which are widespread throughout China, are the gold resources of China and have been extensively used for treating multiple diseases for thousands of years. However, Chinese herbs and herb formulas are characterized by complicated components, resulting in various targets and pathways in treating diseases, which limits their approval and applications. With the development of chemistry and pharmacology, active ingredients of TCM and herbs and underlying mechanisms have been further identified and confirmed by pharmacists and chemists, which improved, to some extent, awkward limitations, approval, and applications regarding TCM and herbs. In this review, we summarized various herbs, herb formulas, natural compounds, and phytochemicals isolated from herbs that have the potential for regulating multiple biological processes via modulation of the JAK/STAT signaling pathway based on the published work. Our study will provide support for revealing TCM, their active compounds that treat diseases, and the underlying mechanism, further improving the rapid spread of TCM to the world.

A Facile Strategy for Development of pH-Sensing Indicator Films Based on Red Cabbage Puree and Polyvinyl Alcohol for Monitoring Fish Freshness.

This study aimed to develop a novel pH-sensing biopolymer film based on red cabbage puree (RCP) incorporated with polyvinyl alcohol (PVA), which was utilized for monitoring fish freshness during storage at 25 °C. A homogenized RCP suspension with a mean particle size of 12.86 ± 0.03 µm and a total anthocyanin concentration of 292.17 ± 2.65 mg/L was directly used as a film-forming substance and anthocyanin source to blend with PVA, showing visual changes in color and ultraviolet-visible spectra within a pH of 2-12. Rheological and microstructural studies certified the strong interactions and good compatibility between the RCP and PVA, resulting in better mechanical properties and water resistance of the composite film than those of a pure RCP film, but without affecting its pH sensitivity. When used for fish freshness monitoring at 25 °C, the developed RCP/PVA film presented visible color differences from purple to yellow, which corresponded to the spoilage threshold of the total volatile basic nitrogen and the total viable count in fish samples. The study highlights that anthocyanin-rich purees of fruits and vegetables, in this case red cabbage puree, can be fully utilized to develop eco-friendly pH-sensing indicator films for intelligent food packaging.

Diaphyseal and Metaphyseal Modeling Defects-Clinical Findings and Identification of WRAP53 Deficiency in Craniometadiaphyseal Dysplasia.

Background: Diaphyseal and metaphyseal modeling defects lead to severe changes in bone mass and shape, which are common features in osteoporosis that linked to non-vertebral fractures. Original mechanism of diaphyseal and metaphyseal modeling defects has proved elusive. Studying rare syndromes can elucidate mechanisms of common disorders and identify potential therapeutic targets. Methods: We evaluated a family pedigree with craniometadiaphyseal dysplasia (CRMDD, OMIM 269300), a genetic disorder that is characterized by cortical-bone thinning, limb deformity, and absent of normal metaphyseal flaring and diaphyseal constriction. Systemic radiographic examination and serum hormone test were made for this rare disease. One patient and her two normal parents were examined by means of whole-exome sequencing (WES) to identify the candidate pathogenic gene and rule out mucopolysaccharidosis and Prader-Willi Syndrome by means of Sanger sequencing. Results: There are several conspicuous radiographic characteristics: (1) bullet-shaped phalanges, (2) long and narrow pelvic inlet, absent of supra-acetabular constriction, (3) round rod-shaped long tubular bones, (4) prominent aiploic mastoid, (5) bending-shaped limb, genua varus and genu varum, and (6) congenital dislocation of elbow. Here, we did not find any wormian bones, and there are several typical clinical characteristics: (1) macrocephaly and wide jaw, (2) Avatar elf-shaped ears, pointed and protruding ears, (3) hypertrophy of limbs, (4) flat feet and giant hand phenomenon, (5) nail dystrophy, (6) limb deformity, (7) high-arched palate, (8) superficial hemangiomas, (9) tall stature, and intellectual disability. In this patient, we found biallelic frameshift deletion mutations in WRAP53, and those two mutations were transmitted from her parents respectively. Conclusions: We describe her clinical and radiological findings and presented a new subtype without wormian bones and with a tall stature. Our study showed that craniometadiaphyseal dysplasia was caused by a deficiency of WRAP53 with autosomal recessive inheritance.

A Multifunctional Tb-MOF Detector for H2O2, Fe3+, Cr2O7 2-, and TPA Explosive Featuring Coexistence of Binuclear and Tetranuclear Clusters.

A novel three-dimensional microporous terbium(III) metal-organic framework (Tb-MOF) named as [Tb10 (DBA)6(OH)4(H2O)5]·(H3O)4 (1), was successfully obtained by a solvothermal method based on terbium nitrate and 5-di(2',4'-dicarboxylphenyl) benzoic acid (H5DBA). The Tb-MOF has been characterized by single crystal X-ray diffraction, elemental analysis, thermogravimetry, and fluorescence properties, and the purity was further confirmed by powder X-ray diffraction (PXRD) analysis. Structural analysis shows that there are two kinds of metal cluster species: binuclear and tetranuclear, which are linked by H5DBA ligands in two µ7 high coordination fashions into a three-dimensional microporous framework. Fluorescence studies show that the Tb-MOF can detect H2O2, Fe3+, and Cr2O7 2- with high sensitivity and selectivity and can also be used for electrochemical detection of exposed 2,4,6-trinitrophenylamine (TPA) in water. The highly selective and sensitive detection ability of the Tb-MOF might make it a potential multifunctional sensor in the future.

Serological investigation of IgG and IgE antibodies against food antigens in patients with inflammatory bowel disease.

BACKGROUND: Food antigens have been shown to participate in the etiopathogenesis of inflammatory bowel disease (IBD), but their clinical value in IBD is still unclear. AIM: To analyze the levels of specific immunoglobulin G (IgG) and E (IgE) antibodies against food antigens in IBD patients and to determine their clinical value in the pathogenesis of IBD. METHODS: We performed a retrospective study based on patients who visited the First Affiliated Hospital of Nanjing Medical University between August 2016 and January 2018. A total of 137 IBD patients, including 40 patients with ulcerative colitis (UC) and 97 patients with Crohn's disease (CD), and 50 healthy controls (HCs), were recruited. Serum food-specific IgG antibodies were detected by semi-quantitative enzyme-linked immunosorbent assay, and serum food-specific IgE antibodies were measured by Western blot. The value of food-specific IgG antibodies was compared among different groups, and potent factors related to these antibodies were explored by binary logistic regression. RESULTS: Food-specific IgG antibodies were detected in 57.5% of UC patients, in 90.72% of CD patients and in 42% of HCs. A significantly high prevalence and titer of food-specific IgG antibodies were observed in CD patients compared to UC patients and HCs. The number of IgG-positive foods was greater in CD and UC patients than in HCs (CD vs HCs, P = 0.000; UC vs HCs, P = 0.029). The top five food antigens that caused positive specific IgG antibodies in CD patients were tomato (80.68%), corn (69.32%), egg (63.64%), rice (61.36%), and soybean (46.59%). The foods that caused positive specific IgG antibodies in UC patients were egg (60.87%), corn (47.83%), tomato (47.83%), rice (26.09%), and soybean (21.74%). Significantly higher levels of total food-specific IgG were detected in IBD patients treated with anti-TNFα therapy compared to patients receiving steroids and immunosuppressants (anti-TNFα vs steroids, P = 0.000; anti-TNFα vs immunosuppressants, P = 0.000; anti-TNFα vs steroids + immunosuppressants, P = 0.003). A decrease in food-specific IgG levels was detected in IBD patients after receiving anti-TNFα therapy (P = 0.007). Patients who smoked and CD patients were prone to developing serum food-specific IgG antibodies [Smoke: OR (95%CI): 17.6 (1.91-162.26), P = 0.011; CD patients: OR (95%CI): 12.48 (3.45-45.09), P = 0.000]. There was no difference in the prevalence of food-specific IgE antibodies among CD patients (57.1%), UC patients (65.2%) and HCs (60%) (P = 0.831). CONCLUSION: CD patients have a higher prevalence of food-specific IgG antibodies than UC patients and HCs. IBD patients are prone to rice, corn, tomato and soybean intolerance. Smoking may be a risk factor in the occurrence of food-specific IgG antibodies. Food-specific IgG antibodies may be a potential method in the diagnosis and management of food intolerance in IBD.

Silencing of LncRNA BDNF-AS attenuates Aß25-35-induced neurotoxicity in PC12 cells by suppressing cell apoptosis and oxidative stress.

OBJECTIVE: To explore the effects of long non-coding RNA (lncRNA) brain-derived neurotrophic factor anti-sense (BDNF-AS) on the Aß25-35-induced neurotoxicity in PC12 cells. METHODS: PC12 cells were induced by Aß25-35 to construct cell injury models of Alzheimer's disease (AD), and then transfected with siRNA-BDNF-AS. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to detect the expressions of BDNF-AS and BDNF. Besides, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Hoechst33342 staining were utilized to analyze the cell viability and apoptosis, respectively, Western blotting to evaluate the protein expressions, immunofluorescence to assess the Cytochrome C (Cyt C) release, and Rhodamine 123 (Rh123) to measure the mitochondrial membrane potential (MMP).The evaluation of oxidative stress was conducted via the determination of the levels of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT). RESULTS: Aß25-35 apparently increased BDNF-AS but decreased BDNF in PC12 cells, which also reduced viability and induced apoptosis of PC12 cells. Silencing of BDNF-AS could significantly up-regulate BDNF in Aß25-35-induced PC12 cells, with the elevated cell viability. Moreover, silencing BDNF-AS inhibited the apoptosis of Aß25-35-induced PC12 cells, suppressed the release of Cyt C, reduced the expression of cleaved caspase-3 and Bax, and lowered the mean fluorescence intensity (MFI) of Rh123, but it elevated the expression of Bcl-2. Besides, silencing BDNF-AS also reduced ROS intensity and MDA content, but enhanced the activities of SOD and CAT. CONCLUSION: Silencing BDNF-AS exerts protective functions to increase the viability, inhibit the apoptosis and oxidative stress of Aß25-35-induced PC12 cells by negative regulation of BDNF. ABBREVIATIONS: Aß25-35: amyloid beta peptide 25-35; AD: Alzheimer's disease; LncRNA BDNF-AS: long non-coding RNA brain-derived neurotrophic factor anti-sense; OS: Oxidative stress.

Phytochemical, cytotoxic and chemotaxonomic study on Ajuga forrestii Diels (Labiatae).

A phytochemical investigation of Ajuga forrestii Diels led to the isolation of 14 compounds, including eight neo-clerodane diterpenes (1-8), two phytoecdysteroids (9, 11), one stigmastane sterol (10) and three iridoid glycosides (12-14). The structures of these compounds were identified by spectroscopic methods and a comparison of their data with those reported in the literature. This is the first report of compounds 1-14 from A. forrestii. The cytotoxic activities of the aqueous extract of A. forrestii and several compounds have been studied and the chemotaxonomic significance of isolated compounds has also been summarised.

Protective efficacy of a single salvianolic acid A treatment on photothrombosis-induced sustained spatial memory impairments.

With respect to the high burden of ischemic stroke and the absence of pharmacological treatment for promoting rehabilitation, promising candidates with specific effects on long-term functional recovery are highly desired. Candidates need reasonable experimental paradigms to evaluate the long-term functional outcome focused on ischemia-induced sensorimotor and memory deficits. "Danshen", a traditional Chinese herb, has long been used to treat coronary and cerebral vascular diseases as well as dementia. Salvianolic acid A (SAA), one of the major active ingredients of Danshen, was demonstrated to be effective in protecting against cerebral ischemic injury. Here, employing an experimental stroke model induced by photothrombosis in the unilateral frontal cortex of rats, we investigated whether SAA has long-term protective effects on ischemia-induced sensorimotor and memory deficits in our behavioral tests. The results indicated that a single SAA treatment improved the cortical ischemia-induced sensorimotor deficits during 15 days' cylinder test period, and alleviated ischemia-induced sustained spatial memory impairments during the 2 months' dependent Morris Water Maze (MWM) tests. In addition, either ischemic injury or SAA treatment did not show any changes compared with sham group in other behavioral tests including rotarod tests, swimming speed in MWM tests, open field tests, elevated plus maze tests, treadmill tests and forced swimming tests. The results reveal that the cognitive deficits are not the results of animal's anxiety or confounding motor impairments. Overall, the present paradigm appears suitable for the preclinical evaluation of the long-term effects of pharmacological treatments on ischemic stroke. Meanwhile, SAA might have therapeutic potential for the treatment of memory deficits associated with ischemic stroke.

Clinical efficacy of entecavir in the treatment of hepatitis B cirrhosis complicated with hepatic diabetes / 中国生化药物杂志

Objective To investigate the clinical efficacy of entecavir in the treatment of hepatitis B cirrhosis associated with hepatic diabetes. Methods 78 cases of hepatitis B Hepatic Cirrhosis with diabetes were selectedfrom June 2015 to December 2016, and randomly divided into study group (n=39) and control group (n=39). The study group were treated with entecavir, the control group were treated with adefovir dipivoxil. Two groups of patients with hepatitis B cirrhosis combined with hepatic diabetes were treated continuously for 48 weeks, the changes of FPG, HOMA-β, beta and HOMA-IR were recorded. Results There was no significant difference between the two groups in the treatment of FPG, HOMA-β, and HOMA-IR. After treatment, the two groups of FPG and HOMA-IR decreased significantly compared with before treatment, while HOMA-β was significantly higher than before treatment, but the changes of the above indexes in the study group were better than those in the control group (P<0.05). Conclusion Entecavir combined with hypoglycemic agents can achieve more ideal blood glucose control effect in patients with hepatitis B cirrhosis combined with hepatic diabetes..

Clinical efficacy of entecavir in the treatment of hepatitis B cirrhosis complicated with hepatic diabetes / 中国生化药物杂志

Objective To investigate the clinical efficacy of entecavir in the treatment of hepatitis B cirrhosis associated with hepatic diabetes. Methods 78 cases of hepatitis B Hepatic Cirrhosis with diabetes were selectedfrom June 2015 to December 2016, and randomly divided into study group (n=39) and control group (n=39). The study group were treated with entecavir, the control group were treated with adefovir dipivoxil. Two groups of patients with hepatitis B cirrhosis combined with hepatic diabetes were treated continuously for 48 weeks, the changes of FPG, HOMA-β, beta and HOMA-IR were recorded. Results There was no significant difference between the two groups in the treatment of FPG, HOMA-β, and HOMA-IR. After treatment, the two groups of FPG and HOMA-IR decreased significantly compared with before treatment, while HOMA-β was significantly higher than before treatment, but the changes of the above indexes in the study group were better than those in the control group (P<0.05). Conclusion Entecavir combined with hypoglycemic agents can achieve more ideal blood glucose control effect in patients with hepatitis B cirrhosis combined with hepatic diabetes..

Novel Insights into the Pathogenesis of Hirschsprung's-associated Enterocolitis.

OBJECTIVE: To systematically summary the updated results about the pathogenesis of Hirschsprung's-associated enterocolitis (HAEC). Besides, we discussed the research key and direction based on these results. DATA SOURCES: Our data cited in this review were obtained mainly from PubMed from 1975 to 2015, with keywords "Hirschsprung enterocolitis", "Hirschsprung's enterocolitis", "Hirschsprung's-associated enterocolitis", "Hirschsprung-associated enterocolitis", "HAEC", and "EC". STUDY SELECTION: Articles regarding the pathogenesis of HAEC were selected, and the articles mainly regarding the diagnosis, surgical approach, treatment, and follow-up were excluded. RESULTS: Several factors, mainly including mucus barrier, intestinal microbiota, and immune function, as well as some other factors such as genetic variations and surgical reasons, have been found to be related to the pathogenesis of HAEC. Changed quantity and barrier property of mucus, different composition of microbiota, and an abnormal immune state work together or separately trigger HAEC. CONCLUSIONS: The maintenance of intestinal homeostasis is due to a well cooperation of microbiota, mucus barrier, and immune system. If any part presents abnormal, intestinal homeostasis will be broken. Meanwhile, for patients with Hirschsprung's disease or HAEC, dysfunction of these parts has been found. Thus, the happening of HAEC may be mainly attributed to the disorders of intestinal microbiota, mucus barrier, and immune system.

3D detection and extraction of bladder tumors via MR virtual cystoscopy.

PURPOSE: This paper proposes a pipeline for the detection and extraction of 3D regions of bladder tumors via MR virtual cystoscopy. METHODS: After the acquisition of volumetric bladder images with a high-resolution T2-weighted 3D sequence, the inner and outer surfaces of the bladder wall were segmented simultaneously by a coupled directional level-set method. Based on the Laplacian method, a potential field was built up between two surfaces so that the thickness of each voxel within the bladder wall was estimated. To detect bladder abnormalities, four volume-based morphological features, including bent rate, shape index, wall thickness, and a novel morphological feature, which reflects bent rate difference between the inner and outer surfaces, were extracted. The combination of these four features was used to detect seeds on the inner surface by using selected filtering criterion. Then all points on streamlines started from detected seeds formed 3D candidate regions. Finally the fuzzy c-means clustering with spatial information (sFCM) was used to extract tumors from surrounding bladder wall tissues in candidate regions. RESULTS: The proposed pipeline was evaluated by a database of MR bladder images acquired from ten patients with bladder cancer. To find an optimal feature combination for tumor detection, the performance of different combinations of these features was evaluated with different filtering criteria. With the combination of all four features, the computer-aided detection pipeline shows a high performance of 100 % sensitivity with 2.3 FPs/case. Comparing with tumor regions delineated by radiological experts, the average overlap ratio of tumor regions extracted by sFCM is 86.3 %. CONCLUSIONS: The experimental result demonstrates the feasibility of the proposed pipeline on the detection and extraction of bladder tumors. It may provide an effective way to achieve the goal of evaluating the whole bladder for tumor detection and local staging.

EMP acupoint stimulation conducive to increase the effect of weight reduction through aerobic exercise.

PURPOSE: to observe whether weight reduction through aerobic exercise is more effective with the intervention of electromagnetic pulse (EMP) Acupoint stimulation. METHOD: based on the need of this study, we choose 48 young and middle-aged obese patients and randomly divide them into Experimental Group and Control Group with each group consisting of 24 of them. The Control Group has an aerobic endurance running each day with each running lasting for about 45 minutes and covering about 6 KM; the Experimental Group has the same exercise as the CG, but, after their running, stimulation by electromagnetic pulse meter is given to their Sanyinjiao Acupoint and Zushanli Acupoint. Then the content of body fat and the lipid indexes of the two groups before the treatment are compared with their counterparts after the said treatment. RESULT: after 6 weeks of treatment, the various indexes of the two groups are improved to different degrees (P<0.05); the effect on lipid indexes of the Experimental Group is obviously better than that of the Control Group (P<0.05), but the content of their body fat shows no conspicuous difference (P>0.05). CONCLUSION: after 6 weeks of treatment, the intervention of EMP acupoint stimulation can more evidently improve the lipid indexes than it has done to the group only having aerobic exercise; but it has little effect in terms of the improvement of the body fat content, which may be because of the short time of such intervention.

Curative effect of Tai Chi exercise in combination with auricular plaster therapy on improving obesity patient with secondary hyperlipidemia.

OBJECTIVE: Observe the effect of Tai Chi in combination with auricular plaster therapy on treating obesity patient with secondary hyperlipidemia. METHOD: Select 45 patients who suffer from simple obesity and secondary hyperlipidemia and then adopt random digital table to divide them into a Tai Chi group, an auricular plaster therapy group and a combination group. Each group consists of 15 patients. The patients in Tai Chi group are trained with Tai Chi twice a day, while those in auricular plaster therapy are treated with auricular plaster therapy 3-5 times a day and those in the combination group are trained with Tai Chi and auricular plaster therapy twice a day. BMI, body fat percentage and blood lipid indexes are respectively detected for the selected patients in the three groups before treatment and after 180 days' treatment. RESULTS: After 180 days' treatment, BMI index and body fat percentage of Tai Chi group are significantly improved in comparison with those before treatment (P<0.05) and the blood lipid index also presents the improvement trend, but the overall effect is not obvious; body fat percentage and BMI index of the auricular plaster therapy group are not improved obviously in comparison with those before the treatment (P>0.05) but the blood lipid index is improved significantly (P<0.05); each index of the combination group is improved significantly compared with those before treatment (P<0.05). By comparing the improvement effect after treatment with that of the other two groups, P<0.05, the difference shows the statistical significance and the treatment effect is more obvious. CONCLUSION: As for the patient suffering from simple obesity and secondary hyperlipidemia, Tai Chi exercise in combination with auricular plaster therapy can show the obvious synergistic therapeutic effect and thus the combined curative effect is obviously superior to that of the single therapy method.

Band offsets of non-polar A-plane GaN/AlN and AlN/GaN heterostructures measured by X-ray photoemission spectroscopy.

The band offsets of non-polar A-plane GaN/AlN and AlN/GaN heterojunctions are measured by X-ray photoemission spectroscopy. A large forward-backward asymmetry is observed in the non-polar GaN/AlN and AlN/GaN heterojunctions. The valence-band offsets in the non-polar A-plane GaN/AlN and AlN/GaN heterojunctions are determined to be 1.33 ± 0.16 and 0.73 ± 0.16 eV, respectively. The large valence-band offset difference of 0.6 eV between the non-polar GaN/AlN and AlN/GaN heterojunctions is considered to be due to piezoelectric strain effect in the non-polar heterojunction overlayers.

The cell death and DNA damages caused by the Tet-On regulating HSV-tk/GCV suicide gene system in MCF-7 cells.

Ganciclovir (GCV) affects the molecular mechanism of cell death and DNA damage by the rAAV (recombinant adeno-associated virus)-mediated Tet-On/HSV-tk/GCV suicide gene system in human breast cancer cell line MCF-7. A rAAV/TRE/Tet-On/HSV-tk combining a Tet-On regulating system and a suicide gene HSV-tk was used to transfect human breast cancer cell line MCF-7, and therapeutic effects on this system were studied. Afterwards, we used RT-PCR, western blotting, and a modified comet-assay to explore the potential mechanism of the HSV-tk/GCV suicide gene system in breast cancer treatments. MTT assay has shown that the cell number of GCV+rAAV+Dox group was significantly decreased compared with that of other groups after treatment and flow cytometric analysis detected that there was a substantial increase of S phase cells in this group, which means the HSV-tk/GCV suicide gene system probably works on the cell cycle. RT-PCR detected the expression level of p21 increased and PCNA had an opposite trend. Western blotting detected the protein expression of p21 and p53 increased and PCNA, CDK1, cyclin B decreased in GCV+rAAV+Dox group. The modified comet-assay shown that the very small extra fragments generated by the GCV+rAAV+Dox group treatment are visible as a small cloud extending from the comet in the direction of electrophoresis. The therapeutic mechanism of the HSV-tk/GCV suicide gene system on human breast cancer cell line MCF-7 is probably by upregulating the expression of p21 through a p53-dependent DNA damage signalling pathway, leading the decrease of protein expression of PCNA, cyclin B, CDK1 in MCF-7 cells and promoting the cell cycle arrest at G1/S phase. In summary, the HSV-tk/GCV suicide gene system arouses the death of MCF-7 cells from blocking the cell cycle and DNA damage.

miR-218 modulate hepatocellular carcinoma cell proliferation through PTEN/AKT/PI3K pathway and HoxA10.

PURPOSE: To investigate the regulatory mechanism of miR-218 in human hepatocellular carcinoma (HCC). METHODS: qPCR was used to compare the expression levels miR-218 among six hepatocellular carcinoma cell lines and normal liver tissues. After transfecting MHCC97L cells with either miR-218 mimics or miR-218 inhibitor, western blotting was used to examine the expressing patterns of cyclinD1, p21, and PTEN/AKT/PI3K signaling pathway-related proteins. MTT and colony forming assay was used to assess the capability of cell proliferation. Bioinformatic method was applied to predict the binding of miR-218 on HoxA10, and western blotting was used to examine the modulatory effect of miR-218 AND HoxA10 on PTEN/AKT/PI3K pathway in HCC. RESULTS: The expression levels of miR-218 were frequently lower in HCC cell lines than in normal liver tissues. Over-expression of miR-218 in HCC cells significantly decreased cell proliferation whereas inhibiting miR-218 promoted cancer cell proliferation. Western blotting analysis demonstrated that tumorigenesis related protein cyclin D1 and p21, as well as PTEN/AKT/PI3K signaling pathways were actively modulated by miR-218 in HCC cells. The expression of endogenous HoxA10 was also down-regulated by miR-218 over-expression, and silencing HoxA10 directly activated PTEN in HCC cells. CONCLUSION: Modulation of miR-218 actively affected HCC cancer cell development. The regulatory mechanism of miR-218 in HCC cells was acting through PTEN/AKT/PI3K pathway and possibly associated with HoxA10.

[Preliminary exploration on submucosal tunneling endoscopic resection for middle and lower esophagus submucosal tumors].

OBJECTIVE: To evaluate the efficacy and safety of submucosal tunneling endoscopic resection (STER) in the treatment of middle and lower esophagus submucosal tumors (SMT) originating from muscularis propria (MP) layer. METHODS: A total number of 33 esophagus submucosal tumor (SMT) originating from MP layer underwent tumor resection by STER after endoscopic ultrasonography (EUS) and CT examination at Endoscopy Center, Department of Gastroenterology, First Affiliated Hospital, Nanjing Medical University from March 2012 to March 2013. There were 17 males and 16 females with an age range of (50 ± 10) years. Their lesion size, lesion origin, pathology, operative duration and complication rate were analyzed. RESULTS: Among them, the origins were of submucosal (n = 4, 12.1%), superficial muscularis propria layer (SMP) (n = 18, 54.6%), deep muscularis layer (DMP) (n = 10, 30.3%) and serosa (n = 1, 3.0%). There were single tumor (n = 30, 90.9%), double tumors (n = 2, 6.1%) and triple tumors (n = 1, 3.0%). Except for 1 case of non-resected hemangioma, 36 operative specimens were examined pathologically, including 30 leiomyomas tumors (83.3%), 5 stromal tumors (GIST) (13.9%) and 1 lipoma tumor (2.8%). Thirty-two lesions were successfully resected by STER with a complete resection rate of 97.0%. Average lesion size was (1.7 ± 1.0) cm and average operative duration (49 ± 26) min. A number of (7.8 ± 2.5) hemostatic clips were used to close the mucosal incision site. Subcutaneous emphysema occurred in 3 patients (9.1%) while puncture and pneumothorax developed in one case (3.0%). All of them recovered uneventfully through conservative treatments. CONCLUSIONS: As a new safe, efficacious and feasible treatment for middle and lower esophagus submucosal tumors, STER may completely resect SMT and provide accurate histopathological evaluations. And it is feasible to regain the mucosal integrity of GI tract and prevent the occurrences of leakage and secondary infections.