Retinal degeneration diseases, including age related macular degeneration, retinitis pigmentosa and glaucoma optic atrophy etc, are characterized by the degeneration of retinal neural cells, retinal photoreceptor cells and retinal pigment epithelium cells, etc.Retinal degeneration diseases are the main cause of blindness. It has been generally assumed that the mature mammalian retinal cell is devoid of repair and regenerative capability, hence it's irreversible of retinal cell apoptosis. Currently, there is no efficient method to regenerate retinal organ. However, stem cell would hold great promise to restore visual function, so as to provide an alternative medical care with the biological property of differentiation and proliferation into target cells to replace degenerated retinal cells. This review focus on the sources of retinal stem cells and their applications on retinal degeneration diseases.

Objective To evaluate neuropsychiatric adverse effects of cycloserine therapy for multidrug resistant pulmonary tuberculosis (MDR-TB).Methods A total of 82 patients with MDR-TB who were enrolled in Global Fund Round Five MDR-TB Control Program were admitted in Center for Diagnosis and Treatment of Tuberculosis,Hangzhou Red Cross Hospital from May to December 2012.All patients received the standard treatment containing cycloserine for MDR-TB.The adverse reactions during the treatment were recorded,and symptom checklist-90 (SCL-90) scores at different time points were compared with t test.Results Adverse reactions were observed in 66 patients (66/82,80.5％) within 3 months after the initial treatment.Common adverse reactions included arthralgia (42.7％),gastrointestinal reactions (40.2％),central nervous system symptoms (22.0％) and electrolytes disturbance (17.1％).Nine patients had severe neuropsychiatric symptoms characterized by convulsions,depression,anxiety,schizophrenia and attempting suicide,6 of whom had used fluoroquinolones before the study.The above symptoms were relieved after stopping cycloserine or antitubercular agents,and cycloserine was replaced in the following treatment.The total SCL-90 score,depression and anxiety scores were significantly higher during onset of symptoms than those one month after the following treatment (t =2.241,2.301 and 5.659,P ＜ 0.05).Conclusion Cycloserine may induce severe neuropsychiatric adverse reactions in patients receiving standard treatment for MDR-TB.

OBJECTIVE: To identify the structure of illegal additive D- (-) -isoascorbic acid in vitamin C Yinqiao tablets and to screen for it in different samples. METHODS: The separation of D- (-) - isoascorbic acid in vitamin C Yinqiao tablets was performed on a CAPCELL PAK NH2 column (4.6 mm × 250 mm, 5 μm) with mobile phase consisting acetonitrile -20 mmol · L-1 ammonium acetate (adjusted to pH 2.5 with formic acid) (60:40), the flow rate was 1.0 mL · min-1. D-(-)-isoascorbic acid was detected by diode array detector at 246 nm. Electrospray ionization (ESI) source in positive ion mode was applied for the identification with following parameters; nebulizer pressure of 220 kPa, drying gas temperature of 350°C, flow rate of 12 L · min-1, and capillary voltage of 3.5 kV. The additive D-(-)- isoascorbic acid in vitamin C Yinqiao tablets was separated and prepared by HPLC for further structure confirmation by 13C-NMR. RESULTS: The structure of the additive was elucidated by analyzing the fragment of [M + H]+ of the additive and comparing with that the reference substance. The additive was further confirmed as D-(-)- isoascorbic acid by 13C-NMR. D-(-) - isoascorbic acid was detected in 64 of 366 batch samples. CONCLUSION: The LC-MS method is accurate and sensitive, which can be used for rapid screening and identification of the illegal additive D- (-) - isoascorbic acid in vitamin C Yinqiao tablets. Copyright 2012 by the Chinese Pharmaceutical Association.

Objective: To evaluate the efficiency of deep-learning based computer aided diagnosis system (DL-CAD) in detecting fractures on DR chest anteroposterior films, and to explore its capability of assisting the junior radiologist. Methods: ①Experiment 1: A total of 547 DR chest anteroposterior films, including 361 patients with 983 chest fractures and 186 without chest fractures were retrospectively analyzed. The predictive performance of DL-CAD for fracture was evaluated. ②Experiment 2: Totally 397 patients were randomly selected from experiment 1, including 211 cases with 604 chest fractures and 186 cases without chest fractures. The results of DL-CAD alone (group 1), a junior radiology resident alone (group 2), a junior radiology resident aided with DL-CAD (group 3) and a senior radiologist alone (group 4) were recorded and compared, respectively. Results: ①For experiment 1: Among 983 fractures, DL-CAD identified 672 fractures, 641 were correctly identified and 31 were misdiagnosed, with a sensitivity of 65.21% (641/983) and F-measure of 77.46%. Out of a total of 361 fracture cases, DL-CAD identified 314 cases, misdiagnosed 6 cases, with a sensitivity of 86.98% (314/361) and F-measure of 92.22%. ②Experiment 2: The sensitivity of fracture detection was 62.09% (375/604), 61.59% (372/604), 86.75% (524/604) and 83.44% (504/604), and the F-measure was 75.38%, 74.62%, 90.74%, 89.84% for group 1, 2, 3 and 4, respectively. The detection efficacy of group 3 and 4 were both higher than that of group 1 and 2 (all P<0.001). There was no significant difference between group 1 and group 2, nor group 3 and group 4 (both P>0.05). Conclusion: DL-CAD software showed good detection effect of fractures on DR chest anteroposterior films, which could effectively improve the diagnostic performance of junior radiologist in fracture detection.

Objective: To investigate the antitumor effect of volatile oil from Sinapis Albae Semen (VOSAS) on H22-bearing mice and to determine the mechanism. Methods: To establish the H22 implanted hepatocellular carcinoma animal model which was used to analyze the effect of VOSAS on the growth of transplanted tumor. Mice were divided into five groups 24 h after modeling: model, cytoxan (CTX, 25 mg/kg) positive control, low-, mid-, and high-dose (20, 40, and 80 mg/kg) VOSAS groups. The mice were ip administered once daily for 10 d. Morphological changes in H22 solid tumor cells were observed by both Hematoxylin-eosin (HE) and acridine orange (AO) staining. The expression of Bax and Bcl-2 in the tumor tissue was determined using immunohistochemistry. Results: VOSAS could inhibit the tumor growth and extend the life span of H22-bearing mice (P < 0.01); and it could also raise the expression of Bax while suppress the expression of Bcl-2; the antitumor effect of VOSAS on H22-bearing mice demonstrated a good dose-effect relationship, but the high-dose group of the volatile oil has obvious toxicity and side effects on the mice. Conclusion: VOSAS could inhibit the growth of H22 tumor cells and the mechanism may be related to up-regulating the expression of Bax and down-regulating the expression of Bcl-2, and the induction of apoptosis.

This study was aimed to quantitatively detect the expression levels of pre-miR-17 and pre-miR-20a in acute leukemia patients and eight kinds of leukemia cell lines, and to investigate the anti-leukemia mechanism of miR-17 and miR-20a silence mediated by miRNA Sponge. Quantitative real-time PCR was used to detect the mRNA expression levels of pre-miR-17 and pre-miR-20a in patients with various types of leukemia and leukemia cell lines. The Jurkat cells over-expressing miR-17 and miR-20a were transfected with recombinant lentivirus-transfecting units targeted at miR-17 and miR-20a plus 6 µg/ml of polybrene. Then the proliferation ability and cell cycle of Jurkat cells was evaluated by CCK-8 and flow cytometry respectively. The results showed that the expression level of pre-miR-17 and pre-miR-20a in all leukemia patients was significantly higher than that in normal group(P < 0.05), the expression of pre-miR-17 and pre-miR-20a in acute lymphoid leukemia was significantly higher than that in acute myeloid leukemia(P < 0.05), and the pre-miR-17 and pre-miR-20a expression level did not correlate significantly with high white blood cell count>20.0×10(9)/L(P > 0.05). The miR-17 and miR-20a silencing mediated by miRNA Sponge led to a significant decrease of cell growth, restored G1 accumulation and increase of cell apoptosis. It is concluded that the expression of miR-17 and miR-20a is upregulated in leukemia patients, which may contribute to leukemogenesis. Over-expressed miR-17 and miR-20a promote cell growth and cell cycle progression, and inhibit apoptosis through negatively-regulating P21 and E2F1 after-transcriptionally.
Cell Line, Tumor ; Gene Silencing ; Genetic Vectors ; Humans ; Leukemia ; genetics ; pathology ; Leukemia, Myeloid, Acute ; genetics ; MicroRNAs ; genetics

This study was aimed to construct lentivirus-mediated shRNA expression vector targeting Bmi-1 and establish a stable cell line U266-li, so as to pave the way for further research on function of Bmi-1 and application of shRNA to gene therapy. One pair of oligonucleotide sequences targeted at human Bmi-1 mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pLVTHM vector. Virus particles were collected after the control or shRNA vectors were co-transfected with the psPAX2 packaging plasmid and the plasmid pMD2.G was enveloped into HEK-293T cells by using Lipofectamine2000. The U266 cells were transduced with 5 × 10(6) recombinant lentivirus-transducing units plus 6 µg/ml of polybrene. Real-time PCR and Western blot were used respectively to detect the expression of Bmi-1 and P14 after lentivirus transduction. DNA sequencing demonstrated that the lentivirus RNAi vector of Bmi-1 was constructed successfully and the virus was packaged in 293T cells. The titer of virus was 5 × 10(7) TU/ml. Stable transfected U266 cell line was established. As was expected, the mRNA and protein levels of Bmi-1 was reduced significantly in U266 cells after lentivirus transduction, whereas the mRNA and protein levels of P14 was upregulated. It is concluded that the lentiviral RNAi vector of Bmi-1 is constructed, and U266 stable cell line is established.
Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Polycomb Repressive Complex 1 ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics

BACKGROUNDAmyloid β (Aβ) has been established as a key factor for the pathological changes in the brains of patients with Alzheimer's disease (AD), and cellular senescence is closely associated with aging and cognitive impairment. However, it remains blurred whether, in the AD brains, Aβ accelerates the neuronal senescence and whether this senescence, in turn, impairs the cognitive function. This study aimed to explore the expression of senescence-associated genes in the hippocampal tissue from young to aged 5XFAD mice and their age-matched wild type (WT) mice to determine whether senescent neurons are present in the transgenic AD mouse model.

METHODSThe 5XFAD mice and age-matched wild type mice, both raised from 1 to 18 months, were enrolled in the study. The senescence-associated genes in the hippocampus were analyzed and differentially expressed genes (DEGs) were screened by quantitative real-time polymerase chain reaction. Cognitive performance of the mice was evaluated by Y-maze and Morris water maze tests. Oligomeric Aβ (oAβ) (1-42) was applied to culture primary neurons to simulate the in vivo manifestation. Aging-related proteins were detected by Western blotting analysis and immunofluorescence.

RESULTSIn 5XFAD mice, of all the DEGs, the senescence-associated marker p16 was most significantly increased, even at the early age. It was mainly localized in neurons, with a marginal expression in astrocytes (labeled as glutamine synthetase), nil expression in activated microglia (labeled as Iba1), and negatively correlated with the spatial cognitive impairments of 5XFAD mice. oAβ (1-42) induced the production of senescence-related protein p16, but not p53 in vitro, which was in line with the in vivo manifestation.

CONCLUSIONSoAβ-accelerated neuronal senescence may be associated with the cognitive impairment in 5XFAD mice. Senescence-associated marker p16 can serve as an indicator to estimate the cognitive prognosis for AD population.

Alzheimer Disease ; metabolism ; physiopathology ; Amyloid Precursor Protein Secretases ; genetics ; metabolism ; Amyloid beta-Peptides ; metabolism ; Amyloid beta-Protein Precursor ; metabolism ; Animals ; Aspartic Acid Endopeptidases ; genetics ; metabolism ; Brain ; metabolism ; physiopathology ; Cells, Cultured ; Cellular Senescence ; genetics ; physiology ; Cognition ; physiology ; Cognition Disorders ; metabolism ; physiopathology ; Disease Models, Animal ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Neurons ; metabolism ; pathology ; Real-Time Polymerase Chain Reaction

OBJECTIVEThis study was aimed to detect the expression of HIF-1α in acute myeloid leukemia (AML) except acute promyelocyte leukemia (APL) and investigate the relationship of its expression levels with clinical parameters and prognosis.

METHODSThe primary AML cells were collected from peripheral blood of 53 newly diagnosed AML patients by using CD3 negative sorting. The expression of HIF-1α was measured by real-time fluorescent quantitative PCR (FQ-PCR) , and the relationship between expression level of HIF-1α and clinical parameters (age, sex, WBC count, clinical typing, prognosis) was analysed according to relative expression level. Furthermore, Western blot was used to detect the protein level of HIF-1α in AML patients with or without extramedullary infiltration.

RESULTSThe expression level of HIF-1α did not correlate with age, sex, WBC count, Hb level, Plt count and the percentage of blast. There was no significant difference of HIF-1α expression between different AML subtype based on FAB. The higher level of HIF-1α was found in AML patients who did not get complete remission after one or two courses of chemotherapy, however, the difference was not statistically significant. The relapse rate was higher in AML patients with the higher expression of HIF-1α. In addition, the higher level of HIF-1α mRNA and protein were found in bone marrow of AML patients with extramedullary infiltration (P < 0.01). The negative correlation between HIF-1α and PTEN was observed (r = -0.48, P = 0.001).

CONCLUSIONSOverexpression of HIF-1α are closely related with extramedullary infiltration and prognosis of acute myeloid leukemia, and may be used as an early indicator of extramedullary infiltration and prognosis.

Granulocyte Precursor Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; Leukemia, Myeloid, Acute ; Prognosis ; RNA, Messenger ; Recurrence ; Remission Induction

OBJECTIVE: To investigate the role of nucleophosmin (NPM) in the proliferation of chronic myeloid leukemia cells (K562 cells) and its mechanism by RNAi technology. METHODS: shRNA was used to inhibit the expression of NPM. The expression of NPM gene was detected by real-time quantitative PCR. The effect of inhibiting NPM gene on cell proliferation was detected by MTS assay. Change of cell cycle was detected by flow cytometry. Western blot was used to detect the expression of cell cycle-related proteins. RESULTS: The shRNA lentiviral vector targeting at NPM gene was successfully constructed and used to transfect the K562 cells. The results showed that compared with the control groups, suppression of NPM gene expression in K562 cells could inhibit the cell proliferation and decrease the cell colony formation. Moreover, interference of NPM gene could prolong G/G phase and arrest cell cycle, which may be related to the down-regulation of NPM gene expression and activation of p21 protein expression, thereby inhibited the formation of CDK2/ Cyclin E complex. CONCLUSION Down-regulation of NPM gene expression in K562 cells can induce cell cycle arrest and inhibit cell proliferation.
Apoptosis ; Cell Proliferation ; Gene Knockdown Techniques ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Nuclear Proteins