OBJECTIVETo establish a hepatic metastatic subline of nasopharyngeal carcinoma (NPC) cell line.

METHODSNPC cells metastatic to the liver were isolated from nude mice and the invasion and metastatic ability of the cells was observed in vivo and in vitro.

RESULTS AND CONCLUSIONThe invasion and metastasis activity of 5-8F-H3B-EGFP (an in vivo isolate with enhanced liver metastatic behaviors) were enhanced obviously in comparison with the parent cell line 5-8F-EGFP. This subline may be useful for cloning genes related to liver metastasis of NPC.

Animals ; Carcinoma, Squamous Cell ; genetics ; metabolism ; secondary ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; Disease Models, Animal ; Female ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Keratins ; analysis ; Liver Neoplasms, Experimental ; genetics ; metabolism ; secondary ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Metastasis ; Neoplasm Transplantation

In this work, a rapid analysis method basing on ultraviolet spectroscopy was established for the determination of danshensu, protocatechuic aldehyde, rosmarinci acid, lithospermic acid and salvianolic acid B in the extraction process of Danhong injection. In the extraction process of Danshen and Honghua crude drugs, 44 extraction solution samples were collected and the contents of the five components were determined by HPLC analysis. The ultraviolet spectra of the samples were collected. Partial least square regression was used to establish the multivariate calibration models between the ultraviolet spectra and the contents of the five components. The results showed that the established models could predict the contents of the five components in the extraction solution accurately. The ultraviolet spectroscopy method established in this work can be used for rapid analysis of the intermediates of Danhong injection, which may be applied for the quality control in the manufacturing process.
Chemistry, Pharmaceutical ; standards ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Quality Control ; Salvia miltiorrhiza ; chemistry ; Spectrophotometry, Ultraviolet ; methods

OBJECTIVETo analyze the gene expression profiles of transcription factor-related genes in nasopharyngeal carcinoma (NPC) tissues and normal nasopharyngeal tissues using a cDNA microarray membrane for exploring the regulatory mechanism of differential gene express in NPC tissues.

METHODSThe total RNAs from 24 NPC tissues and 24 pooled normal nasopharyngeal tissues were reverse transcribed and labeled with alpha-(32)P-dCTP. The resultant cDNAs were hybridized to GF211 microarray, and the signals were analyzed by Pathway 4.0 software. RT-PCR was carried out to confirm the results.

RESULTSAmong the 1625 differentially expressed genes detected in NPC and nasopharyngeal tissues, 35 transcription factor-related genes were identified with either up- or down-regulation.

CONCLUSIONThese differentially expressed transcription factor-related genes in NPC tissues might play a role in the regulation of NPC-related gene expression.

Carcinoma, Squamous Cell ; genetics ; pathology ; E2F1 Transcription Factor ; genetics ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Nuclear Proteins ; genetics ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics ; Tumor Cells, Cultured

OBJECTIVETo investigate the clinical efficacy and safety of modified stapled transanal rectal resection (STARR) combined with perioperative pelvic floor biofeedback therapy (POPFBFT) in treating obstructed defecation syndrome (ODS).

METHODSThirty female ODS patients underwent modified STARR (resection and suture was performed in rectocele with one staple) combined with POPFBFT in Department of Colorectal and Anal Surgery, The First Hospital of Jilin university from October 2013 to March 2015. Before the modified STARR, patients received a course of POPFBFT (20 min/time, 2 times/d, 10 times as a course), and another 2 courses were carried out in clinic after discharge. Efficacy evaluation included general conditions of patients, morbidity of postoperative complication, overall subjective satisfaction (excellent: without any symptoms; good: 1 to 2 times of laxatives per month and without the need of any other auxiliary defecation; fairly good: more than 3 times of laxatives per month ; poor: with no improvement; excellent, good, fairly good are defined as effective), Longo ODS score (range 0 to 40 points, the higher the score, the more severe the symptoms), gastrointestinal quality of life index(GIQLI)(range 0 to 144 points, the lower the score, the more severe the symptoms), anorectal manometry and defecography examination. The follow-up lasted 12 months after operation (ended at April 2016).

RESULTSAverage age of 30 patients was 57(46 to 72) years and Longo ODS score of every patient was ≥9 before operation. The modified STARR was completed successfully in all the 30 patients with average operation time of 25 (18 to 34) min and average hospital stay of 6(4 to 9) d. Postoperative complications included pain(20%, 6/30), urinary retention (16.7%, 5/30), anorectal heaviness (6.7%, 2/30), and fecal urgency(26.7%, 8/30). Anaorectal heaviness and fecal urgency disappeared within 3 months. No severe complications, such as postoperative bleeding, infection, rectovaginal fistula, anastomotic dehiscence and anal incontinence were observed. The effective rate of overall subjective satisfaction was 93.3%(28/30) during the follow-up of 12 months. There was no significant difference in Longo ODS score between pre- POPFBFT and pre-operation (pre- POPFBFT: 32.95±3.22, pre-operation: 32.85±3.62, t=1.472, P=0.163). Compared with pre-POPFBFT, Longo ODS score at 1 week after operation decreased (t=4.306, P=0.000), moreover, score at 1 month after operation was lower than that at 1 week (13.05±7.49 vs. 15.00±7.17, t=7.322, P=0.000), while no significant differences were found among 1, 3, 6, 12 months after operation (F=2.111, P=0.107). Likewise, there was no significant difference in GIQLI score between pre-POPFBFT and pre-operation (pre-POPFBFT: 79.39±17.14, pre-operation: 76.65±17.56, t=1.735, P=0.096). Compared with the pre-POPFBFT, GIQLI score at 1 week after operation increased (t=4.714, P=0.000), moreover, GIQLI score at 1 month after operation was higher than that at 1 week (102.26±19.24 vs 91.31±21.35, t=5.628, P=0.000), while no significant differences were found among 1, 3, 6, 12 months after operation(F=1.211, P=0.313). In comparison with pre- POPFBFT, parameters of defecography examination at 12 months after operation showed obvious improvement: the rectocele decreased from (34.1±0.4) mm to (3.1±0.3) mm (t=6.847, P=0.000), anorectal angle during defecation increased from (123.8±6.7)degree to (134.7±8.5)degree, enlargement of anorectal angle during defecation increased from (29.1±3.5)degree to (37.1±5.3)degree, while no significant differences in descend of perineum, anorectal angles at rest as well as parameters of anorectal manometry were found (all P>0.05).

CONCLUSIONModified STARR combined with POPFBFT is safe and effective for ODS patients.

Aged ; Anal Canal ; surgery ; Biofeedback, Psychology ; physiology ; Constipation ; rehabilitation ; surgery ; Defecation ; Defecography ; Digestive System Surgical Procedures ; methods ; rehabilitation ; Female ; Humans ; Length of Stay ; Middle Aged ; Operative Time ; Pain, Postoperative ; etiology ; Pelvic Floor ; physiology ; Postoperative Complications ; Quality of Life ; Rectocele ; Surgical Stapling ; Suture Techniques ; Treatment Outcome ; Urinary Retention ; etiology

Objective:Through analysis of the development status of pediatric and the problems faced by pediatric young scientists, combined with the situation of research management, this paper puts forward some suggestions.Methods:Collect the identified problems of pediatric development, summarize the common ones and propose methodological strategies for scientific research management.Results:The development of pediatrics faces problems such as insufficient resources, unreasonable resource allocation and talents shortage.Conclusions:It is recommended to support the development of pediatric according to optimize the scientific research management model, establish a reasonable evaluation and incentive mechanism, establish a pediatric talent training model and team, improve the pediatric research level, and obtain policy support for salary, talent introduction and training, promotion of professional titles, research resources and so on.

Background: The risk and benefit of tissue plasminogen activator (tPA) for aged>80 years with acute ischemic stroke (AIS) are controversial. In this study, we investigated the safety and efficacy of tPA in this population and utilized the artificial neural network (ANN) to established outcome predictive models. Methods: We retrospectively reviewed the stroke registry data of patients with AIS, aged >80 years who arrived at the hospital within 3 hours from the onset of symptoms. The characteristics and the outcomes, presented as modified Rankin Scale (mRS), and mortality rate at 3 months between the tPA-treated and non-tPA groups were analyzed. An ANN algorithm was applied to establish predictive models. Results: A total of 80 patients aged>80 years with AIS were identified, and 49 of them received tPA. After adequate training, our ANN models accurately predicted the outcomes with the area under the receiver operating characteristic curves of 0.974, and a low error to predict the mRS score at 3 months. After applying our prediction model to those in the non-tPA group, we demonstrated the potential benefits in those patients if they had undergone tPA therapy. Conclusions: Our results show that ANN can be a potentially useful tool for predicting the treatment outcomes of tPA. Such novel machine learning-based models may help with therapeutic decision making in clinical settings.

PURPOSE: Whether tamoxifen affects the risk of neurodegenerative disease is controversial. This nationwide population-based study investigated the risk of Parkinson's disease (PD) associated with tamoxifen treatment in female patients with breast cancer using Taiwan's National Health Insurance Research Database. METHODS: A total of 5,185 and 5,592 female patients with breast cancer who did and did not, respectively, receive tamoxifen treatment between 2000 and 2009 were included in the study. Patients who subsequently developed PD were identified. A Cox proportional hazards model was used to compare the risk of PD between the aforementioned groups. RESULTS: Tamoxifen did not significantly increase the crude rate of developing PD in female patients with breast cancer (tamoxifen group, 16/5,169; non-tamoxifen group, 11/5,581; p=0.246). Tamoxifen did not significantly increase the adjusted hazard ratio (aHR) for subsequently developing PD (aHR, 1.310; 95% confidence interval [CI], 0.605–2.837; p=0.494). However, tamoxifen significantly increased the risk of PD among patients followed up for more than 6 years (aHR, 2.435; 95% CI, 1.008–5.882; p=0.048). CONCLUSION: Tamoxifen treatment may increase the risk of PD in Taiwanese female patients with breast cancer more than 6 years after the initiation of treatment.
Asian Continental Ancestry Group* ; Breast Neoplasms* ; Breast* ; Female* ; Humans ; National Health Programs ; Neurodegenerative Diseases ; Parkinson Disease* ; Proportional Hazards Models ; Tamoxifen*

BACKGROUNDCytomegalovirus (CMV) retinitis is the most severe intraocular complication that results in total retinal destruction and loss of visual acuity in patients with acquired immunodeficiency syndrome (AIDS). This study aimed to investigate the fundus characteristics, systemic manifestations and therapeutic outcomes of CMV retinitis associated with AIDS.

METHODSIt was a retrospective case series. CMV retinitis was present in 39 eyes (25 patients). Best corrected visual acuities, anterior segment, fundus features, fundus fluorescence angiography (FFA) and CD4(+) T-lymphocyte counts of the patients with CMV retinitis associated with AIDS were analyzed. Intravitreal injections of ganciclovir (400 µg) were performed in 4 eyes (2 patients).

RESULTSRetinal vasculitis, dense, full-thickness, yellow-white lesions along vascular distribution with irregular granules at the border, and hemorrhage on the retinal surface were present in 28 eyes. The vitreous was clear or mildly opaque. Late stage of the retinopathy was demonstrated in 8 eyes characterized as atrophic retina, sclerotic and attenuated vessels, retinal pigment epithelium (RPE) atrophy, and optic nerve atrophy. Retinal detachment was found in 3 eyes. The average CD4(+) T-lymphocyte count in peripheral blood of the patients with CMV retinitis was (30.6 ± 25.3) × 10(6)/L (range, (0 - 85) × 10(6)/L). After intravitreal injections of ganciclovir, visual acuity was improved and fundus lesions regressed.

CONCLUSIONSCMV retinitis is the most severe and the most common intraocular complication in patients with AIDS. For the patients with yellow-white retinal lesions, hemorrhage and retinal vasculitis without clear cause, human immunodeficiency virus (HIV) serology should be performed. Routine eye examination is also indicated in HIV positive patients.

Acquired Immunodeficiency Syndrome ; complications ; immunology ; metabolism ; Adult ; Antiviral Agents ; pharmacology ; CD4-Positive T-Lymphocytes ; metabolism ; Cytomegalovirus Retinitis ; drug therapy ; etiology ; immunology ; metabolism ; Female ; Fluorescein Angiography ; Ganciclovir ; pharmacology ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo examine the expression patterns of short palate, lung and nasal epithelium clone 1 (SPLUNC1) gene in human tissues.

METHODSIn situ hybridization was used to detect the expression of SPLUNC1 gene in 37 different human tissues.

RESULTSWe found that SPLUNC1 gene was not expressed in squamous epithelial cells of the palate, epidermis, esophagus, or the esophagus-cardia junction, metaplastic squamous cells in the nasopharynx, trachea, or uterus cervix, or tumor cells of esophageal squamous cell carcinoma or lung squamous cell carcinoma. SPLUNC1 gene was not expressed in the single layer columnar epithelia cells in the stomach, gallbladder, jejunum, colon, endometrium, or uterus cervix. SPLUNC1 expression was detected mainly in pseudostratified columnar epithelial cells in the nasopharynx, trachea and bronchi, and was gradually down-regulated from the upper to lower end of the respiratory tract, but was not detected in the lung tissues. SPLUNC1 expression was detected not only in the duct and serous gland cells in the parotid and submandibular glands, but also in cells of submucosal serous glands in the nasopharynx and lung, but not in the cells of the mucosal glands. The parietal cells of the gastric submucosa and epithelial cells of the lobula and ducts of the mammary glands expressed SPLUNC1. The adenocarcinoma cells in the lung, stomach, colon, mammary gland, uterus endometrium and cervix showed strong expressions of SPLUNC1 gene.

CONCLUSIONSPLUNC1 expression is highly cell-specific in association with the cell functions.

Epithelial Cells ; metabolism ; Gene Expression ; Glycoproteins ; genetics ; metabolism ; Humans ; Organ Specificity ; Phosphoproteins ; genetics ; metabolism

Human tumor necrosis factor alpha (hTNF-alpha) is one of the most important inflammatory cytokines that acts as a mediator in inflammatory and immune response and plays a key role in host defense against infection. The over expression of hTNF-alpha is associated with serious consequences, such as shock, hypotension, thrombus, septicemia and even death. It has been implicated in many autoimmune and inflammatory diseases, such as rheumatoid arthritis, Crohn's disease, chronic heart failure and septic shock. Inhibiting the bio-activity of hTNF-alpha is one of the strategy for the treatment of these diseases. Compared with traditional recombinant protein drugs, small molecule drugs have many advantages, such as high affinity, low immunogenecity and low cost. Systematic evolution of ligands by exponential enrichment (SELEX) is a powerful method for the selection of oligonucleotides that bind with high affinity and specificity to target proteins. Such oligonucleotides are called aptamers, and are potential therapeutics for blocking the activity of pathologically relevant proteins. To obtain oligonucleotide aptamers specifically binding to TNF, a 40nt random DNA combinatorial library flanked by 31nt fixed sequences was chemically synthesized. The random library was amplified with PCR and subjected to selection by SELEX protocol against hTNFalpha. After incubation of the library with hTNFalpha, the mixture was blotted onto Immobilon-NC transfer membrane. The no-specific binding was washed away and the hTNFa binding aptamers were eluted and detached from the target protein. The eluted oligo nucleotides were amplified with PCR and served as the DNA library for the next round selection. After 12 rounds of such selection, the selected aptamers were cloned to pGEM-T vector. Positive clones were identified by restriction enzyme digestion and DNA sequencing. Oligo DNA were synthesized according to the sequence data and tested for their activities. Binding activity of the aptamers to hTNFalpha were detected by ELISA and dot blot with biotin-streptavidin-horseradish peroxidase system. Mouse L929 cells were used to test the anti-hTNFa activity of the DNA aptamers. The aptamers were incubated with hTNFalpha and added to the L929 cells. The results were read under microscope and with MTT staining. It was shown that these DNA aptamers bound to hTNFalpha with high affinity, and can inhibit the cytotoxicity of hTNFalpha on cell culture. The affinity of these aptamers are different and may related to their structure. These ssDNA aptamers are potential for the treatment and diagnosis of hTNFalpha related diseases.
Animals ; Aptamers, Nucleotide ; chemical synthesis ; chemistry ; genetics ; pharmacology ; Biotin ; chemistry ; Cell Line ; Cell Survival ; drug effects ; Enzyme-Linked Immunosorbent Assay ; Gene Library ; Humans ; Mice ; Protein Binding ; SELEX Aptamer Technique ; methods ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; metabolism ; toxicity