Live birth in woman with premature ovarian insufficiency and 46, XY karyotype after chemotherapy and bone marrow transplant: a case report.

BACKGROUND: Premature ovarian insufficiency (POI) is a clinical syndrome defined by loss of ovarian function before the age of 40 years, characterized by elevated serum gonadotropin levels and decreased estrogen levels with menstrual disturbance. POI can be natural or iatrogenic such as after chemotherapy, radiotherapy and surgery. CASE PRESENTATION: In this study, we describe a successful live birth in a 31-year-old woman with POI and 46, XY Karyotype after being treated with chemotherapy and bone marrow transplant (BMT) for acute non-lymphocytic leukemia when she was 17 years old. With amenorrhea or oligomenorrhea for 11 years, her serum level of FSH was up to 35.0 IU/L and 53.0 IU/L taken 4 weeks apart, which can be diagnosed as POI. After controlled ovarian stimulation treatment for three cycles with different protocols and frozen-thawed embryo transfer (FET), she finally got a successful pregnancy and had a live birth later. CONCLUSIONS: This case report serves as a reminder that karyotype of peripheral blood may mislead the diagnosis as disorders of sex development (DSD). It also demonstrates that it is possible for a woman with chemotherapy and bone marrow transplant induced POI can have successful pregnancy and live birth with appropriate therapy. Furthermore, as age may plays a predominant role in fertility rather than residual ovarian reserve, active treatment may be concerned for women with POI at younger age.

Repurpose Open Data to Discover Therapeutics for COVID-19 Using Deep Learning.

There have been more than 2.2 million confirmed cases and over 120 000 deaths from the human coronavirus disease 2019 (COVID-19) pandemic, caused by the novel severe acute respiratory syndrome coronavirus (SARS-CoV-2), in the United States alone. However, there is currently a lack of proven effective medications against COVID-19. Drug repurposing offers a promising route for the development of prevention and treatment strategies for COVID-19. This study reports an integrative, network-based deep-learning methodology to identify repurposable drugs for COVID-19 (termed CoV-KGE). Specifically, we built a comprehensive knowledge graph that includes 15 million edges across 39 types of relationships connecting drugs, diseases, proteins/genes, pathways, and expression from a large scientific corpus of 24 million PubMed publications. Using Amazon's AWS computing resources and a network-based, deep-learning framework, we identified 41 repurposable drugs (including dexamethasone, indomethacin, niclosamide, and toremifene) whose therapeutic associations with COVID-19 were validated by transcriptomic and proteomics data in SARS-CoV-2-infected human cells and data from ongoing clinical trials. Whereas this study by no means recommends specific drugs, it demonstrates a powerful deep-learning methodology to prioritize existing drugs for further investigation, which holds the potential to accelerate therapeutic development for COVID-19.

Increased locomotor activity and non-selective attention and impaired learning ability in SD rats after lentiviral vector-mediated RNA interference of Homer 1a in the brain.

Our previous studies found that Homer 1a, a scaffolding protein localized at the post-synaptic density (PSD) of glutamatergic excitatory synapses, is significantly down-regulated in the brain of spontaneous hypertensive rats (SHR), an animal model of attention deficit hyperactivity disorder (ADHD). Furthermore, a first-line treatment drug for ADHD, methylphenidate, can up-regulate the expression of Homer 1a. To investigate the possible role of Homer 1a in the etiology and pathogenesis of ADHD, a lentiviral vector containing miRNA specific for Homer 1a was constructed in this study. Intracerebroventricular injection of this vector into the brain of Sprague Dawley (SD) rats significantly decreased Homer 1a mRNA and protein expression levels. Compared to their negative controls, these rats displayed a range of abnormal behaviors, including increased locomotor activity and non-selective attention and impaired learning ability. Our results indicated that Homer 1a down-regulation results in deficits in control over behavioral output and learning similar to ADHD.

Rice miR168a-5p regulates seed length, nitrogen allocation and salt tolerance by targeting OsOFP3, OsNPF2.4 and OsAGO1a, respectively.

Rice microRNA168a (osa-miR168a) plays important roles in mediating flowering time, grain yield and vigor, seeding growth, and immunity by targeting the RNA-induced silencing complex component Argonaute 1 (AGO1). However, the functions of miR168a exerted by targeting other genes require further clarification before it could be used in rice molecular breeding. In this study, we identified a new target gene of osa-miR168a-5p (miR168a-5p) in rice called OsOFP3 (ovate family protein 3) and investigated the roles of miR168a-5p in response to brassinosteroids (BRs), salt stress, and nitrogen allocation. Up- and downregulated miR168a-5p expression respectively decreased and increased the expression of the BR-negative regulator OsOPF3. The results of RNA ligase-mediated rapid amplification of cDNA ends (5'RLM-RACE) revealed cleavage sites in OsOPF3 and OsNPF2.4 mRNAs. The phenotype of miR168a-5p transgenic rice was BR-associated and included the lamina bending response to BR, short seeds, and low 1000-grain weight. MicroRNA 168a-5p also regulated the expression of the nitrate transporter, OsNPF2.4, which affected nitrogen allocation, and regulated OsAGO1a expression in response to salt stress. Taken together, rice miR168a-5p regulates BR-associated pathways, nitrogen transport, and stress by targeting OsOFP3, OsNPF2.4, and OsAGO1a, respectively, resulting in a series of important agronomic traits for rice breeding.

Preparation of nitrogen-doped carbon dots and their enhancement on lettuce yield and quality.

Nanotechnology is an effective way to stimulate the yield potential of crops. Various nano-fertilizers and nano-carriers are gradually being developed to bring about a technological revolution in the agricultural industry. As a biocompatible water-soluble nanomaterial, carbon dots (CDs) have attracted the attention of researchers for applications in agriculture. In this study, we prepared nitrogen-doped CDs (N-CDs) as a type of water-soluble carbon nanofertilizer by a one-pot hydrothermal method, and investigated its effects on lettuce biomass and quality. 100 and 200 mg L-1 of N-CDs substantially promoted lettuce biomass accumulation (41.70%), elevated lettuce nutrient content, as well as promoted the accumulation of major nutrients. Moreover, 100 mg L-1 N-CDs increased the chlorophyll a content by 12.68%, significantly increased the electron transport rate (ETR) by 38.61%, significantly increased the light energy conversion efficiency (Y(II)) by 31.24% and increased the Rubisco activity by 60.61%, which are important reasons for its increase in actual photosynthesis rate. N-CDs also have a positive effect on plant nitrogen metabolism by promoting the activity of glutamine synthetase. The significant benefits of N-CDs on lettuce make them have great potential for agricultural yield increase and quality improvement.

Immunoregulatory Activity of Herbal Tea-Derived Carbon Dots.

The activation of immune cells by immunoregulatory active substances can improve the body immunity. Carbon dots (CDs) with immunoregulatory activity are rarely reported. In this study, transmission electron microscopy results demonstrate the existence of CDs in herbal tea, while Fourier transform infrared and X-ray photoelectron spectroscopy results suggest the participation of polyphenol in herbal tea CD (H-CD) formation. The photoluminescence spectrum has shown that H-CDs have fluorescence emission at 565 nm and exhibit an excitation-dependent property. The toxicity and immunostimulatory activity of H-CDs on mouse macrophage RAW264.7 suggested that H-CDs had no toxicity to RAW264.7 cells. Meanwhile, compared with herbal tea, H-CDs have more obvious effect of promoting the expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase. In addition, the secretion of nitric oxide (NO) was promoted by H-CDs. This work suggests that H-CDs have stronger immunoregulatory function than that of original herbal tea, which provides a direction for the application of phenolic hydroxyl-modified CDs in the biomedical field.

Glycine assists in efficient synthesis of herbal carbon dots with enhanced yield and performance.

As a special type of biomass, herbal medicine often contains a variety of biologically active substances, and taking it as a carbon source, it is expected to produce various types of biologically functional carbon dots (CDs). However, there are few reports in this field, especially in achieving enhanced performance of CDs by improving the utilization efficiency of active substances in medicinal materials. In this work, by adding glycine as an auxiliary agent in the preparation of CDs from herbal medicine (Exocarpium Citri Grandis), the carboxyl and amino groups of the adjuvant provided more reactive sites, which greatly improved the yield of CDs (about 6 times). More importantly, the antioxidant and biological activities of herbal CDs were also improved. By controlling the functional groups of adjuvants, the effects of carboxyl and amino groups in adjuvants on the synthesis of herbal CDs were compared. The results reveal that both carboxyl and amino groups can react with the substances in the carbon source, and the influence of amino groups was greater. After adding glycine, the size of the CDs became larger, resulting from the more abundant functional groups on the carbon skeleton, which was the main reason for the improved performance of the CDs. Finally, the biological activity experiment demonstrated that CDs derived from Exocarpium Citri Grandis and glycine could greatly enhance the vitality of cells and activate immune cells, which are expected to be applied in the field of cell reproduction and biological immunity. The method proposed in this work provides a potential strategy for high-yield preparation of CDs from biomass.

Protective effects of Huang Qi Huai granules on adriamycin nephrosis in rats.

Huang Qi Huai (HQH) granules, a mixture of Chinese herbs, contains trametes robiniophila murr, wolfberry fruit, and Polygonatum. We investigated the mechanism of the protective effects of HQH on adriamycin nephrosis (ADR) in rats. Adriamycin nephrotic rats were induced by a single dose of 5 mg/kg adriamycin. For the HQH-treated adriamycin nephrosis group, 1 day after treatment with 5 mg/kg adriamycin, the rats were administered once-daily oral gavage of 2 mg/kg HQH for 15 days. All the rats were killed at day 15. Histological changes were observed by light microscopy and transmission electron microscope. Nephrin and podocin expression levels were measured by real-time RT-PCR and Western blot. Proteinuria was measured by the Bradford protein assay. Serum TNF-α and IL-1ß levels were evaluated by ELISA. Macrophage infiltration was detected by immunohistochemistry and immunoblotting, respectively. ADR rats showed heavy proteinuria, podocyte and tubulointerstitial injury, macrophage infiltration, and increased levels of serum cytokines TNF-α and IL-1ß. HQH significantly ameliorated the adriamycin-induced renal injury. These data were validated in the cultured podocytes. The podocytes were treated by adriamycin in the presence or absence of HQH and nephrin and podocin expression and TNF-α and IL-1ß synthesis and secretion were determined by real-time RT-PCR, immunoblotting, and ELISA, respectively. Adriamycin significantly reduced nephrin and podocin expression, which was significantly restored by the treatment of HQH. HQH treatment inhibited adriamycin-induced TNF-α and IL-1ß expression. Our findings suggest that HQH significantly reduces proteinuria, prevents podocyte injury, and ameliorates tubulointerstitial damage. Inhibition of inflammatory cytokine expression and macrophage infiltration may be the protective mechanism of HQH.

Dose-Dependent Effect of ZnO Quantum Dots for Lettuce Growth.

As the cadmium-free semiconductor quantum dots, ZnO quantum dots (ZnO QDs) have wide potential applications in agriculture. However, the effects of ZnO quantum dots on crop growth and nutritional quality have not been fully studied. In this work, the lettuce was sprayed with different concentrations of ZnO QDs from 50 to 500 mg·L-1 to evaluate their influence on lettuce antioxidant, biomass, and nutritional quality. The results showed that ZnO QDs existed in the lettuce in the form of Zn2+. Lettuce treated with 500 mg·L-1 ZnO QDs would produce a large amount of reactive oxygen species (ROS), which adversely affected the absorption of nutrients, soluble protein content, and chlorophyll content, thus reducing plant biomass. When the concentrations range from 50 to 200 mg·L-1, the antioxidant enzyme systems of lettuce were triggered to counteract the damage caused by excessive ROS. Moreover, ZnO QDs at this level promoted Ca, Mg, Fe, Mn, Zn, and B absorption and accumulation; increased soluble sugar content; and improved the lettuce biomass and nutritional quality.

Construction of Carbon Dots with Color-Tunable Aggregation-Induced Emission by Nitrogen-Induced Intramolecular Charge Transfer.

As one of the most promising fluorescent nanomaterials, the fluorescence of carbon dots (CDs) in solution is extensively studied. Nevertheless, the synthesis of multicolor solid-state fluorescence (SSF) CDs is rarely reported. Herein, CDs with multicolor aggregation-induced emission are prepared using amine molecules, all of them exhibiting dual fluorescence emission at 480 nm (Em-1) and 580-620 nm (Em-2), which is related to the SS bonds of dithiosalicylic acid and the conjugated structure attached to CO/CN bonds, respectively. As a strong electron-withdrawing group, the increase of CN content makes dual-fluorescent groups on the surface of CDs produce push and pull electrons, which determines intramolecular charge transfer (ICT) between the double emission. With the increase in CN content from 35.6% to 58.4%, the ICT efficiency increases from 8.71% to 45.94%, changing the fluorescence of CDs from green to red. The increase of ICT efficiency causes fluorescence quantum yield enhancement by nearly five times and redshift of the fluorescence peak. Finally, based on the multicolor luminescence properties induced by the aggregation of CDs, pattern encryption and white-LED devices are realized. Based on the fat solubility and strong ultraviolet absorption characteristics of CDs, fingerprint detection and leaf anti-UV hazards are applied.

Carbon dots as light converter for plant photosynthesis: Augmenting light coverage and quantum yield effect.

Carbon dots (CDs) with gradient-changed quantum yield (QY) were prepared by regulating the graphitic N and hydroxyl group contents. Then, the QY effect of CDs on plant photosynthesis was studied using chloroplasts and rice plants. After incubation for 2 h in the dark, CDs entered into the chloroplasts and converted ultraviolet radiation to photosynthetically active radiation. By this mechanism, CD1:0.2 (300 µg·mL-1) with a moderate QY of 46.42% significantly increased the photosynthetic activity of chloroplast (200 µg·mL-1) to reduce DCPIP and ferricyanide by 43.77% and 25.45%, respectively. After spraying on rice seedlings, CD1:0.2 (300 µg·mL-1) was evenly distributed in the leaves and resulted in maximum increases in the electron transport rate and photosynthetic efficiency of photosystem II by 29.81% and 29.88%, respectively. Furthermore, CD1:0.2 significantly increased the chlorophyll content and RuBisCO carboxylase activity of rice by 64.53% and 23.39%, respectively. Consequently, significant increases were observed in the growth of CD1:0.2-treated rice, including 18.99%, 64.31%, and 61.79% increases in shoot length, dry weights of shoot and root. These findings contribute to the exploitation of solar energy and agricultural production using CDs in the future.

TiO2/Chlorophyll S-Scheme Composite Photocatalyst with Improved Photocatalytic Bactericidal Performance.

Step-scheme (S-scheme) photocatalysts have been proposed for highly efficient charge separation and strong redox activity in the photocatalysis field. Here, we reported a facile strategy to obtain the S-scheme heterojunction composite TiO2/chlorophyll (Chl). The S-scheme heterojunction enables the significant improvement of electron transfer efficiency at the interfacial heterojunction of TiO2/Chl. Also, the lifted conduction band and valence band of TiO2/Chl resulted in more than 1.61 times generation of reactive oxidizing species, compared to that of bare TiO2. In addition, TiO2/Chl was applied as a photocatalytic bactericidal material to fabricate commercial masks for prolonged life span of the mask. The TiO2/Chl-coated mask filter exhibited excellent bactericidal effect on Escherichia coli under light illumination (2.94 × 107 cfu E. coli were killed by 1 cm-2 coated mask filters within illumination of 3 h), while commercial mask filters showed no bactericidal effect. After three circulation-sterilization tests, the TiO2/Chl-made mask filter maintained the initial bactericidal effect, which greatly extended the life span of the mask that presents a promising strategy to alleviate the supply stress of masks.

Gene expression profiles in the prefrontal cortex of SHR rats by cDNA microarrays.

We have undertaken cDNA microarrays to identify differentially expressed genes in the prefrontal cortex (PFC) of spontaneously hypertensive-rat (SHR), a rodent model of attention deficit hyperactivity disorder (ADHD) versus control Wistar-Kyoto (WKY) rats. The analysis of the gene expression profiles indicated that 57 genes were up-regulated and 97 genes were down-regulated in the PFC of SHR. These predominately expressed genes included genes involved in neural development, immunity, transcription factor, monoamine neurotransmitter, metabolism, signal transduction, apoptosis and so on. Although more detailed analyses are necessary, it is anticipated that further study of genes identified will provide insights into their specific roles in the etiology of ADHD.

Enhanced Photogenerated Electron Transfer in a Semiartificial Photosynthesis System Based on Highly Dispersed Titanium Oxide Nanoparticles.

In this study, a hybrid semiartificial photosynthesis system based on chloroplast (CLP) and titanium oxide nanoparticles (TiO2 NPs) was constructed. 2,6-Dichlorophenolindophenol (DCPIP) reduction by TiO2/CLP complex and methylene blue (MB) reduction by TiO2 were used to determine enhanced photogenerated electron transfer in this hybrid system. The DCPIP reduction by the TiO2/CLP complex showed the same trend as MB reduction by TiO2 as a function of concentration of TiO2 NPs, indicating interception of photogenerated electrons in TiO2 by CLP that leads to enhanced photosynthesis efficiency. Decreased photoluminescence intensity and shortened excited-state lifetime of the TiO2/CLP complex compared to that of pure TiO2 also support electron transfer from TiO2 to CLP. Longer visible light absorption wavelength and increasing valence band edges reveal the narrower band gap of TiO2/CLP, which finally results in the enhanced electron transfer from TiO2 to CLP. Higher ferricyanide reduction and enhanced ATP formation with the TiO2/CLP complex demonstrate the accelerated electron-transfer rate of the electron-transfer chain. This study reveals the mechanism of how TiO2 interacts with CLP to enhance the photosynthesis via constructing a semiartificial photosynthesis system.

Vitamin D supplementation prior to in vitro fertilisation in women with polycystic ovary syndrome: a protocol of a multicentre randomised, double-blind, placebo-controlled clinical trial.

INTRODUCTION: Polycystic ovary syndrome (PCOS) is one of the leading causes of female infertility, affecting around 5% of women of childbearing age in China. Vitamin D insufficiency is common in women with PCOS and is associated with lower live birth rates. However, evidence regarding the effectiveness of vitamin D supplementation in women with PCOS is inconclusive. This multicentre randomised, double-blinded, placebo-controlled trial aims to evaluate the effectiveness of vitamin D supplementation prior to in vitro fertilisation (IVF) on the live birth rate in women with PCOS. METHODS AND ANALYSIS: We plan to enrol women with PCOS scheduled for IVF. After informed consent, eligible participants will be randomised in a 1:1 ratio to receive oral capsules of 4000 IU vitamin D per day or placebo for around 12 weeks until the day of triggering. All IVF procedures will be carried out routinely in each centre. The primary outcome is live birth after the first embryo transfer. The primary analysis will be by intention-to-treat analysis. To demonstrate or refute that treatment with vitamin D results in a 10% higher live birth rate than treatment with placebo, we need to recruit 860 women (48% vs 38% difference, anticipating 10% loss to follow-up and non-compliance, significance level 0.05 and power 80%). ETHICS AND DISSEMINATION: This study has been approved by the Ethics Committee in Women's Hospital of Zhejiang University on 2 March 2020 (reference number: IRB-20200035-R). All participants will provide written informed consent before randomisation. The results of the study will be submitted to scientific conferences and a peer-reviewed journal. TRIAL REGISTRATION NUMBER: NCT04082650.

[Value of CD, MPO, Ki-67 and C-MYC Positive Rate in the Pathological Tissues and C-MYC Gene of Patients with T-LBL/ALL for Predicting Prognosis].

OBJECTIVE: To explore the value of of CD, MPO, Ki-67, C-MYC positive rates in the pathological tissues and C-MYC gene of patients with T-LBL/ALL for predicting Prognosis. METHODS: Ninty cases of T-LBL/ALL patients in our hospital were selected and included in the T-LBL/ALL group, and 30 cases of lymphnode reactive hyperplasia were selected as control group. Immunohistochemical staining was used to detect the changes of CD, MPO, Ki-67 and C-MYC positive rate in 2 groups, and the changes of C-MYC gene were detected by fluorescence in situ hybridization. RESULTS: In 90 patients with T-LBL/ALL, there were CD1a+ 34 cases (37.8%), CD3+ 67 cases (74.4%), epsilon CD3+ 47 cases (52.2%), CD7+ 85 cases (94.4%), CD10+ 33 cases (36.7%), CD34+ 22 cases (24.4%), CD43+ 48 cases (53.3%), CD45RO+ 46 cases (51.1%), CD99+ 88 cases (97.8%), TDT+ 85 cases (94.4%); and CD23, CD20, and MPO all were negative; Ki-67>80% 47 cases (52.2% cases), Ki-67≤80%, 43 cases (47.8%). In 90 T-LBL/ALL patients, the positive rate of C-MYC (66.7%) was significantly higher than the control group (positive rate 0.0%) (P< 0.05); the Ki-67 index, mediastinal widening of T-LBL/ALL patients and the positive rate of C-MYC positively were correlated (P< 0.05). The overall survival rate (44.0%) of C-MYC negative patients was significantly higher than that of C-MYC positive patients (0.0%). The overall survival rate of C-MYC negative patients was significantly higher than that of C-MYC positive patients (P< 0.05).Ann Arbor staging, LDH, bone marrow involvement, mediastinal widening, Ki-67 positive index, and C-MYC protein expression of patients with T-LBL/ALL did not correlated with increased C-MYC gene breakage and copy number (P> 0.05). CONCLUSION: The overall survival rate of C-MYC positive patients decreases, which positively correlates with Ki-67 positive index and mediastinal width, suggesting that the prognosis of the patients with C-MYC protein expression is poorer.

A resistin binding peptide selected by phage display inhibits 3T3-L1 preadipocyte differentiation.

BACKGROUND: Resistin, a newly discovered cysteine-rich hormone secreted mainly by adipose tissues, has been proposed to form a biochemical link between obesity and type 2 diabetes. However, the resistin receptor has not yet been identified. This study aimed to identify resistin binding proteins/receptor. METHODS: Three cDNA fragments with the same 11 bp 5' sequence were found by screening a cDNA phage display library of rat multiple tissues. As the reading frames of the same 11 bp 5' sequence were interrupted by a TGA stop codon, plaque lift assay was consequently used to prove the readthrough phenomenon. The stop codon in the same 11 bp 5' sequence was replaced by tryptophan, and the binding activity of the coded peptide [AWIL, which was designated as resistin binding peptide (RBP)] with resistin was identified by the confocal microscopy technique and the affinity chromatography experiment. pDual GC-resistin and pDual GC-resistin binding peptide were co-transfected into 3T3-L1 cells to confirm the function of resistin binding peptide. RESULTS: Three cDNA fragments with the same 11 bp 5' sequence were found. The TGA stop codon in reading frames of the same 11 bp 5' sequence was proved to be readthroughed. The binding activity of RBP with resistin was consequently identified. The expression of the resistin binding peptide in 3T3-L1 preadipocytes expressing pDual GC-resistin significantly inhibited the adipogenic differentiation. CONCLUSION: RBP could effectively rescue the promoted differentiation of resistin overexpressed 3T3-L1 preadipocyte.

[Transdifferentiation and collagen-synthesis effects of exogenous connective tissue growth factor on renal tubular epithelial cells in vitro].

OBJECTIVE: To investigate the effects of recombinal human connective tissue growth factor (rhCTGF) stimulation on epithelial-myofibroblast transdifferentiation (EMT) and collagen-synthesis in human renal tubular epithelial cell line (HK2) in vitro. METHODS: The cultured HK2 cells were stimulated with rhCTGF of 5 ng/mL. The morphological changes were observed under an inverted microscope. The cells were collected at 0, 3, 6, 12, 24 and 48 hrs after rhCTGF stimulation. The expression of E-cadherin,alpha-smooth muscle actin (alpha-SMA), collagen Ialpha1 (Col Ialpha1) and collagen IValpha1 (Col IValpha1) mRNAs were detected by RT-PCR. RESULTS: rhCTGF stimulation changed the HK2 cell appearance from oval to fusiformdown-regulated the E-cadherin mRNA expression and up-regulated alpha-SMA mRNA expression, but had no effects the Col Ialpha1 and Col IValpha1 mRNA expression. CONCLUSIONS: Exogenous CTGF can mediate the EMT but has no collagen-synthesis effects on HK2 cells.

[Inhibition effects of par-4 antisense oligodeoxynucleotide on apoptosis of PC12 cell induced by glutamate is mediated by ERK1/2].

OBJECTIVE: To investigate the inhibition effects of par-4 antisense oligodeoxynucleotide on apoptosis of PC12 cell induced by glutamate and its signal transduction mechanism. METHODS: (1) Cationic lipid-mediated par-4 antisense oligodeoxynucleotide (Par-4-AS-ODN) was transfected into PC12 cells before they were treated with glutamate. Mismatch oligodeoxynucleotide (MS-ODN) were also transfected into cells as controls. (2) Morphological observation and the detection of anti-apoptosis effects of par-4-AS-ODN on PC12 cells were done with the Laser Scanning confocal Microscope by double staining the cells with acridine orange/ethidium bromide (AO/EB), addition to with flow cytometry. (3) Western blot was used to detect the protein levels of par-4 and phosphorylated ERK(1/2) at threonine-202 and Tyrosine-204. RESULTS: (1) Par-4-AS-ODN significantly suppressed up-regulation of the par-4 protein levels induced by glutamate in PC12 cells. (2) Par-4-AS-ODN could resist the decrease of phosphorylated ERK(1/2) (Thr202/Tyr204) induced by glutamate in PC12 cells. (3) Par-4 AS-ODN could inhibit apoptosis of PC12 cells induced by glutamate. But its inhibition effect could be eliminated by PD98059, a selective MEK(1) inhibitor which could inhibit phosphorylation of ERK(1/2). CONCLUSION: Par-4 AS-ODN may inhibit apoptosis of PC12 cells induced by glutamate, and its inhibition effects may be medicated by the activation of ERK(1/2).

[Suppressive effects of par-4 antisense oligodeoxynucleotide on up-regulation of intracellular calcium concentration in PC12 cell induced by glutamate and its anti-apoptosis effects].

OBJECTIVE: To investigate the uppressive effects of par-4 antisense oligodeoxynucleotide on the up-regulation of intracellular calcium concentration in PC12 cell induced by glutamate and its anti-apoptosis effects. METHODS: Cationic lipid-mediated par-4 antisense oligodeoxynucleotide (par-4-AS-ODN) was transfected into PC12 cells and followed by glutamate for treatment. Mismatch oligodeoxy-nucleotide (MS-ODN) was used as the control. Morphological assessment and evaluation of the anti-apoptosis effects of par-4-AS-ODN on PC12 cells were performed by laser scanning confocal microscopy by double staining of the cells with Hoechest 33258/propidium iodide (Hoe/PI) and flow cytometry respectively. The mRNA and protein levels of calpain 10 and par-4 were measured by RT-PCR and Western blot. Intracellular calcium concentration was determined by using laser scanning confocal microscope with Fura-2/AM as the fluorescent dye. RESULTS: Par-4-AS-ODN repress the increase of par-4 protein in PC12 cell (52.3 +/- 5.0 vs 90.0 +/- 3.2, < 0.01). Par-4-AS-ODN significantly inhibited the apoptosis of PC12 cells induced by glutamate (53% vs 31%, < 0.01). Par-4-AS-ODN significantly suppress the up-regulation of intracellular calcium concentration in PC12 cells induced by glutamate (Rate of fluorescent density: 167.9 +/- 32.4 vs 228.8 +/- 36.8, < 0.01). Par-4-AS-ODN inhibited the increase of calpain 10 mRNA in PC12 cells induced by glutamate (46.3 +/- 3.7 vs 34.8 +/- 2.1, < 0.01). CONCLUSIONS: par-4-AS-ODN enables to inhibit apoptosis of PC12 cells induced by glutamate. The mechanism of the inhibition may be closely related to suppression of the up-regulation of intracellular calcium concentration and calpain transcription expression.