Objective To observe the effects of embedding implantation of collagen and Tetramethylpyrazine in acupoints and electro-acupuncture on the expression of HSP70 in hippocampus of rat with focal cerebral ischemia reperfusion. Methods SD rats were randomly divided into normal group, sham operation group, model group, electro-acupuncture group, embedding medicine in acupoints group. Model of local ischemia was made by middle cerebral artery occlusion, preparation of implantation collagen and electro acupuncture and Tetramethylpyrazine was embedded in the acupoints of Dazhui and Neiguan two-sided. The expression of HSP-70 protein in hippocampus of different time point (24, 72, 120 h) was assayed by immunohistochemistry. Results There were only few positive cells of HSP-70 in normal group and sham operation group. The expression of HSP-70 in each therapy group were all heightened, and achieved the peak 24 h later and then declined in brain tissue after focal cerebral ischemiareperfusion, the differences were relatively prominent compared with the same phase model group (P

OBJECTIVETo evaluate the molecular mechanism of proliferation and invasion inhibition in oral squamous cell carcinoma transfected with recombinant adenovirus-p53 (Ad-p53).

METHODSTca8113 cell lines were transfected with Ad-p53. Then the effect of p53 overexpression on cancer cells proliferation and invasion was observed. The expression of mitogen-activated protein kinase (MAPK), serine/threonine kinase (AKT) signaling pathway related proteins, cell cycle and apoptosis related proteins Cyclin D1, P21 and Bcl-2 were detected by Western blotting analysis.

RESULTSAfter transfected with Ad-p53, the proliferation and invasion of Tca8113 cells were significantly inhibited (P < 0.01) and the apoptosis of Tca8113 cells significantly increased (P < 0.001). The results of Western blotting demonstrated that the protein expression of P53 and P21 significantly increased, Cyclin D1 and Bcl-2 protein expression and phosphorylation of AKT protein significantly decreased (P < 0.01).

CONCLUSIONThe AKT signaling pathway may be a key molecular mechanism for proliferation and invasion inhibition of oral squamous cell carcinoma caused by p53. The protein of Cyclin D1, P21 and Bcl-2 may be the downstream targets of AKT signaling pathway. This may provide a new evidence for AKT pathway and downstream targets as a promising therapeutic target for malignant tumors.

Apoptosis ; Carcinoma, Squamous Cell ; Cell Cycle ; Cell Proliferation ; Cyclin D1 ; Genes, p53 ; Humans ; Mitogen-Activated Protein Kinases ; Mouth Neoplasms ; Protein-Serine-Threonine Kinases ; Serine ; Signal Transduction ; Transfection

OBJECTIVE: To establish the droplet digital PCR (ddPCR) assay for the detection of NPM1 type A mutation in patients with acute myeloid leukemia (AML), and to evaluate its specificity, sensitivity and its value in clinical application. METHODS: NPM1 mutant and wildtype plasmids were used to verify the performance of ddPCR. Both ddPCR and Sanger sequencing were used to detect the bone marrow samples of 87 AML patients, which were confirmed by next generation sequencing (NGS). Moreover, NPM1 mutation burden was dynamically monitored in five patients by ddPCR. RESULTS: The limit of blank (LOB) of ddPCR established for NPM1 mutation detection was 1.1 copies/μl, and the limit of detection (LOD) was 2.43 copies/μl, which had good linearity. Among the 87 newly diagnosed AML patients, ddPCR identified seventeen cases positive for NPM1 mutation (19.5%)， which was consistent with Sanger sequencing. NGS confirmed 12 positive cases, including 8 of type A mutations, 2 of type D mutations, and 2 of rare type mutations. The results of dynamic monitoring of NPM1 mutation burden in 5 patients showed that the NPM1 mutation burden decreased obviously even close to 0, when patients achieve complete remission after chemotherapy. However, the mutation burden was increased again at the time of relapse. CONCLUSION In this study, we established a ddPCR method for detection of NPM1 mutation with good sensitivity and repeatability, which can be used for screening NPM1 mutation in newly diagnosed AML patients and for minimal residual disease monitoring after remission in positive AML patients to guide treatment.
Humans ; Leukemia, Myeloid, Acute/therapy* ; Mutation ; Nuclear Proteins/genetics* ; Nucleophosmin ; Polymerase Chain Reaction ; Prognosis

We isolated a new strain of endophytic Pseudomonas G5 from the stems of Chinese parsley (Coriandrum sativum L.), and it is tentatively identified as Pseudomonas aurantiaca according to analysis of the entire substrate utilization profiles using BIOLOG Microstation system (BIOLOG, Inc, Hayward CA). An array of evidence established that many Gram-negative bacteria employ Quorum sensing (QS) system to regulate gene expression in response to cell density using small diffusible signal molecules, N-acyl homoserine lactones (AHLs), and control diverse phenotypic traits in plant-associated bacteria. In this study, we showed that Pseudomonas sp. strain G5 can produce several types of AHLs at a detectable level using Thin Layer Chromatography (TLC) analysis combined with bioreporter Chromobacterium violaceum CV026 bioassay, and N-hexanoyl-homoserine lactone (HHL, C6-HSL) with Rf value 0.4 is the major signal molecule. Furthermore, we have identified its quorum sensing system composed of PhzI and PhzR by cloning and sequencing of phzI-phzR. PhzI is responsible for synthesis of AHLs signal molecules, and PhzR is a transcriptional regulator. Finally, we heterologously expressed the recombinant plasmid pMD-phzIR in Escherichia coli JM109 and verified it using C. violaceum CV026 bioassay. The phylogenetic analysis using MEGA4 revealed highly similarities exist among the phzIR homologs, suggesting it is evolutionary well conserved in the genus Pseudomonas.
4-Butyrolactone ; analogs & derivatives ; metabolism ; Acyl-Butyrolactones ; metabolism ; Amino Acid Sequence ; Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation, Bacterial ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Phylogeny ; Pseudomonas ; classification ; genetics ; isolation & purification ; Trans-Activators ; biosynthesis ; genetics

Clinical data of a child with acquired immunodeficiency syndrome characterized by ischemic stroke who was admitted to the Pediatric Intensive Care Unit of Children′s Hospital Affiliated to Soochow University in January 2019 were retrospectively analyzed.The child is a 6 years and 4 months old boy with a history of thrombocytopenic purpura and recurrent respiratory infections.The main complaint was " the right limb weakness for more than 10 days" . The head magnetic resonance imaging (MRI) revealed extensive abnormal signals in the bilateral frontal and parietal lobes and the formation of softening foci in the left thalamus and outer capsule.Blood routine showed white blood cell 4.88×10 9/L, lymphocyte ratio 0.291, lymphocyte count 1.42×10 9/L, hemoglobin 99 g/L, and platelet 23×10 9/L.Lymphocyte subsets included CD3 + 84.1%, CD3 + CD4 + 0.2%, CD3 + CD8 + 61.4%, CD4 + /CD8 + 0, CD3 -CD 19+ 9.2%, CD3 -CD 16+ 56+ 6.1%, and CD 19+ CD 23+ 5.8%.Pretransfusion tests suggested human immunodeficiency virus (HIV) (+ ), and that other results were negative.Both parents of the child were infected with HIV.This paper demonstrates that neurological involvement is not rare in HIV infection, and stroke is the most common cause of clinical focal neurological deficits in HIV-infected children.Screening with MRI is recommended for high-risk children with neurologic symptoms or neurocognitive dysfunction.

BACKGROUND:Chitosan biological materials can induce bone marrow mesenchymal stem cells to differentiate toward neurons. As a derivative of chitosan, carboxymethyl chitosan has a series of excelent properties. However, whether carboxymethyl chitosan can induce the neuronal differentiation of bone marrow mesenchymal stem cells remains unclear.OBJECTIVE:To investigate the effect of carboxymethyl chitosan thermosensitive hydrogel on the differentiation of bone marrow mesenchymal stem cells into neurons and the possible mechanism.METHODS:Passage 3 bone marrow mesenchymal stem cells from rats were selected and cultured in carboxymethyl chitosan thermosensitive hydrogel extracts in different concentrations (0, 50, 100, 150, 200, 500 g/L). Control cells were cultured in culture medium with no addition of carboxymethyl chitosan thermosensitive hydrogel extracts. MTT assay was performed to investigate the effects of different concentrations of carboxymethyl chitosan thermosensitive hydrogel extracts on bone marrow mesenchymal stem cell proliferation. Western blot assay was used to explore the effect of 150 g/L carboxymethyl chitosan thermosensitive hydrogel extracts on the expression of neuron-specific enolase, Nestin, Vimentin, NF-M, microtubule associated protein 2, glial fibrillary acidic protein, β3-tubulin, Notch1 and Jag1 protein.RESULTS AND CONCLUSION:MTT assay showed that carboxymethyl chitosan thermosensitive hydrogel promoted the cell proliferation, and the proliferation rate reached the peak at the concentration of 150 g/L. Western blot assay showed that the cells induced by 150 g/L carboxymethyl chitosan thermosensitive hydrogel extract had significant increases in neuron-specific enolase, Nestin, Vimentin, NF-M, microtubule associated protein 2, glial fibrillary acidic protein, and β3-tubulin protein expression, and obvious decreases in Notch1 and Jag1 protein expression in comparison with the control group. These results indicate that the carboxymethyl chitosan thermosensitive hydrogel induces rat bone marrow mesenchymal stem cells to differentiate toward neurons, and suppresses the activity of Notch signal pathway in the process of differentiation.

OBJECTIVETo determine the encapsulation efficiency of silybin liposomes.

METHODAfter Silybin liposomes solution was separated through sphadex G-100 column, eluent collected was determined by the first derivative UV spectrometry.

RESULTThe calibration curve of Silybin was linear in the range of 0.5-30.0 mg x L(-1) (r = 0.9998). The average recovery was 101.7%. The relative standard deviation was 1.8%. The results showed that the encapsulation efficiency ranged from 65.1%-83.0% when the drug/phospholipid ratio varied from 0.02-0.06. The concentration of SLB in liposomes prepared by optimum technique was 760.4 mg x L(-1) (n = 4).

CONCLUSIONThe method is accurate, simple and suitable for the quality control of Silybin liposomes.

Drug Delivery Systems ; Liposomes ; chemistry ; Milk Thistle ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Silymarin ; administration & dosage ; analysis ; isolation & purification ; Spectrophotometry, Ultraviolet ; Technology, Pharmaceutical ; methods

Objective To improve the accuracy of diagnosis and differential diagnosis by summarizing imaging characteristics of spinal giant cell tumor of bone (GCTB).Methods Total 28 cases of spinal GCT were confirmed by pathology,of which 2 localized in cervical vertebra,8 in thoracic vertebra,8 in lumbar vertebra,and the other 10 lesions in the sacrum.All patients underwent X-ray,CT,and MRI scanning.Results (1)The incidence of spinal GCTB in the sacrum is highest,up to 35.7%.Lesions can locate in single or more vertebral bodies.All of the 10 cases with primary lesion locating the sacrum were involved in more vertebral bodies. (2)X-ray and CT showed central or eccentric vertebral destruction with 22 cases involving in adnexal bones,21 cases with bone crest or bony septum and 26 cases with soft tissue mass.(3)Lesions examined with MRI showed inhomogeneous isointense or slight hy-pointense on T1 WI,inhomogeneous hyperintense or isointense on T2 WI and inhomogeneous hyperintense or slight hyperintense on STIR.(4)Expansive bone destruction and soft tissue mass occurred again in postoperative recurrent lesions.Conclusion X-ray,CT and MRI are of significant value in the diagnosis of the spinal GCT.The combination of the three is helpful to improve the accuracy of diagnosis and differential diagnosis.

The invertebrate parvovirus Bombyx mori Densonucleosis Virus type 3(Zhenjiang isolate), named BmDNV-3, is a kind of bidensovirus. The most obvious characteristic in the genome of BmDNV-3 is that it has 2 sets of DNA molecular (VD1, VD2),and each of them is encapsidated respectively in the form of single-stranded liner DNA ( + VD1, - VD1, + VD2, - VD2) in equal percentage. So the BmDNV-3 has 4 kinds of virions. Furthermore the sequence of BmDNV-3 is able to encode DNA polymerase itself. Some strains of silkworm revealed complete resistance to BmDNV-3, so they didn' t fall sick. To investigate the difference in the process of infection and replication between the 2 virions ( VD1, VD2) of this bidensovirus, and the difference of the increment in the resistant or susceptible host, the 5th instar larvae of the susceptible silkworm strain (HUABA 35) and the resistant silkworm strain(QIUFENG d) were inoculated determinate dose of BmDNV-3 by oral ingestion. Then the midgut were collected at 9 timepoints. The silkworm cytoplasm actin A3 was used to be normalized gene, so the number of cells in collected tissue could be determined. The result shows that whatever in the susceptible silkworm strain or in the resistant one, the copies of VD1 and VD2 in the genome of BmDNV-3 collected at the different timepoint were almost at the equal level respectively, so that the VD1 and VD2 were replicated with synchronization. The process of infection in the susceptible silkworm strain was devided into 3 partitions, latent period( 2 - 12 hours post inoculation), exponential phase (12 - 36 hours post inoculation)and stationary phase (36- 96 hours post inoculation and there are about 2 x 10(5) copies per cell) . In the resistant silkworm strain, the virus were replicated at a very low level, that was from 6 - 10 copies 2 hours post inoculation to 150 - 200 copies 96 hours post inoculation (about 20 times) . So we predict that the resistance in some of the silkworm strains from BmDNV-3 was a kind of chronic representation that the host carried virus without being caused flacherie.
Animals ; Bombyx ; genetics ; virology ; DNA, Viral ; genetics ; Densovirus ; genetics ; physiology ; Female ; Genome, Viral ; genetics ; Host-Pathogen Interactions ; Male ; Polymerase Chain Reaction ; methods ; Time Factors ; Virion ; genetics ; physiology ; Virus Replication ; genetics

OBJECTIVETo investigate the mechanism of norcantharidin (NCTD)-induced SMMC-7721 hepatoma cell apoptosis.

METHODSSMMC-7721 cell growth inhibition was measured by the MTT method. Apoptosis was detected by Annexin V/propidium iodide staining. The mitochondrial membrane potential was measured by flow cytometry. Western blot analysis was used to evaluate the level of cytochrome c, caspase-3, AIF, Bcl-2 and Bax expression.

RESULTSNCTD inhibited SMMC-7721 cell growth in a time- and dose-dependent manner. The cells treated with NCTD showed the loss of mitochondrial membrane potential. The activities of caspase-3, cytochrome c, AIF, and Bax were up-regulated after NCTD treatment at different doses. The expression of Bcl-2 was decreased after treatment with NCTD.

CONCLUSIONSNCTD could induce SMMC-7721 cell apoptosis. The activation of the mitochondrial pathway was involved in the process of NCTD-induced SMMC-7721 cell apoptosis.

Apoptosis ; drug effects ; Apoptosis Inducing Factor ; metabolism ; Blotting, Western ; Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cytochromes c ; metabolism ; Flow Cytometry ; Humans ; Liver Neoplasms ; enzymology ; metabolism ; pathology ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects ; metabolism ; bcl-2-Associated X Protein ; metabolism