Objective To investigate the association between the expression of heparanase(Hpa) and the invasion of choriocarcinoma by studying the expression of Hpa in human choriocarcinoma cell lines JEG-3 and JAR and human chorionic villous tissues.Methods(1)Matrigcl invasion assays were used to detect in vitro invasive ability of JEG-3 cells and JAR cells.(2)Expression of Hpa protein in the human chorionic villous tissues and choriocarcinoma cell lines(JEG-3 cells and JAR cells)were detected by immunocytochemistry and western blot.Results(1)The invasive cell number was significantly larger in JEG-3 cells than in JAR cells(191?17 vs 106?13,P

Objective To explore the significance of case managers using WeChat platform in the treatment process and extended care for patients with laryngeal cancer.Methods From November 2014 to May 2015, 30 patients with partial resection or total resection of laryngeal cancer in the department of head and neck surgery were selected as a research subjects.Case managers provided fully supports,such as establishing WeChat group,replying online consultations,popularizing health education knowledge,providing psychological counselings.Psychological assessment of the patients was conducted by social support scale(SSRS)and anxiety scale(SAS)in the beginning of case management.Results comparison was performed before and after three months intervention.Results The score of subjective support of patients before intervention was (25.13 ±4.81)points,which after intervention was (28.33 ± 4.41)points.The score of support availability before intervention was (7.20 ±1.99)points,which after intervention was (35.27 ±5.90)points.The differences were statistically significant(t =-2.685,-5.280,2.917,all P <0.05).Conclusion The WeChat platform mediated by the case management is effective in the management of the psychological health of patients with laryngeal cancer,which can effectively promote the rehabilitation of patients.

Objective: To explore the mechanism of acupuncture in regulating cognitive deficits in insomnia rats by observing the effect of acupuncture on microglia in thalamic reticular nucleus (TRN). Methods: Thirty rats were randomly divided into a control group, a model group and an acupuncture group, with 10 rats in each group. The insomnia model was established by intraperitoneal injection of para-chlorophenylalanine (PCPA) once a day for 2 d. Rats in the control group were intraperitoneally injected with the same amount of normal saline. Rats in the acupuncture group received acupuncture at Neiguan (PC 6) and Zusanli (ST 36) for 5 consecutive days. The CLOCKLAB 2 data acquisition system was used to dynamically observe the sleep of the rats throughout the experiment. The cognition of rats was evaluated by event-related potentials (ERPs). After intervention, brain tissue was extracted. Immunofluorescence was used to test the fluorescence expression in TRN region. The concentrations of interleukin (IL)-1β and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay. Results: After intraperitoneal injection of PCPA suspension, the spontaneous activity in light period of rats in the model group and acupuncture group increased significantly compared with the control group (both P<0.01). After acupuncture treatment, the rats in the acupuncture group had much less spontaneous activity during the light period than those in the model group (P<0.01), and the results indicated that acupuncture could effectively improve the sleep quality of insomnia rats. Compared with the control group, rats in the model group showed that the P3 latency, the average optical density of microglia, and the concentrations of IL-1β and TNF-α increased significantly (all P<0.05), and the P3 amplitude decreased significantly (P<0.01). Compared with the model group, rats in the acupuncture group presented that the P3 latency, the average optical density of microglia, and the concentrations of IL-1β and TNF-α were significantly decreased (all P<0.05), and the amplitude of P3 was significantly increased (P<0.05). Conclusion: Acupuncture possesses an ability to improve the cognitive state in insomnia rats. The mechanism may be related to inhibiting the microglial activation, diminishing the levels of pro-inflammatory mediators like IL-1β and TNF-α, and promoting the recovery of central nervous system function.

The artificial seeds of Dendrobium huoshanese was produced with Auxiliary buds, Protocorm-like bodies, and adventitious shoots. By using orthogonal experiment, we studied the effect of Maltose (%), hormone rate between 6-BA (mgx L(-1)) and NAA (mgL(-1)), active carbon (%), sodium alginate (%), time of ion exchange (min) on germination rate of artificial seeds of D. huoshanese. Then the leaking rate of maltose and variation of pH value of artificial seed capsule during vegetating of artificial seeds of D. huoshanese was measured. The results show that maltose played the most important role in inducing D. huoshanese artificial seeds to germinate. The optimal combination was: maltose 4%, hormone rate between 6-BA (mg x L(-1)) and NAA (mg x L(-1)) 10:1, active carbon 0.1%, sodium Alginate 4%, time of ion exchange is 10 min. Protocorm-like bodies were appropriate propagartor, the germination rate of artificial seeds of D. huoshanese takeing Protocorm-like bodies as the propagartors is 90.1%. After germination, the survival rate of seedlings of artificial seeds was 80.6%, the leaking rate of maltose of artificial seed capsule was 0.52%, and the pH value of artificial seed capsule decreased during the process of vegetation of artificial seeds. After having been stored at 4 degrees C for 20 d, the germination rate of artificial seeds of D. huoshanese takeing Auxiliary buds, Protocorm-like bodies, Adventitious shoots as the propagartors were 3.3%, 10.6%, 5.2%. Under natural conditions the germination rate was 13.8% after 10.0 g/L carbendazim was appended into artificial seed capsule. This result provides a foundation of manufacture and further study of the artificial seeds of D. huoshanese.
Alginates ; pharmacology ; Charcoal ; pharmacology ; Culture Techniques ; methods ; Dendrobium ; growth & development ; Germination ; Glucuronic Acid ; pharmacology ; Hexuronic Acids ; pharmacology ; Maltose ; pharmacology ; Plant Shoots ; growth & development ; Seeds ; growth & development

To investigate the effect of recombinant human growth hormone (rhGH) on JAK2-STAT3 pathway and the growth of gastric cancer cell lines at different GHR expression status, the eukaryotic expression vector targeting human GHR (pGPU6/GFP/Neo-shGHR and pGPU6/GFP/Neo-scramble) was constructed and transfected into MGC803 cells by Lipofectamine 2000. Stable expressive cell lines were obtained by G418 screening. The expression of GHR was analyzed by Western blotting. After being stimulated with rhGH, cell growth was detected by MTT assay. Cell cycle and apoptosis were examined by flow cytometry. The components of JAK2/STAT3 signaling pathway were detected by Western blotting. There is no significant difference of GHR expression between MGC803 and pGPU6/GFP/Neo-scramble-transfected cells (named as MGC803-NC) (P > 0.05). Compared with MGC803, the GHR expression in pGPU6/GFP/Neo-shGHR-transfected cells (named as MGC803-shGHR) decreased significantly (protein decreased 50%). The cells were treated with rhGH at 0, 150 and 300 ng x mL(-1), the growth rate of MGC803 and MGC803-NC increased significantly, PI and the number of G2/M phase cells all increased significantly, and apoptosis decreased significantly. Western blotting revealed that the expression of pJAK2 and pSTAT3 was up-regulated after being treated with rhGH in MGC803 and MGC803-NC cells. In contrast, similar change was not observed in MGC803-shGHR cells. Knockdown of GHR gene may decrease the sensitivity of gastric cancer cells to rhGH, and down-regulating of components of the expression of JAK2/STAT3 signaling pathway may be the potential mechanisms.
Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Human Growth Hormone ; genetics ; pharmacology ; Humans ; Janus Kinase 2 ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Somatotropin ; genetics ; metabolism ; Recombinant Proteins ; genetics ; pharmacology ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology ; Transfection

Osteoclasts are multinucleated giant cell, which derived from mononuclear myeloid hematopoietic stem cells with the function of bone absorption. Osteoclasts plays a key role in bone metabolism, therefore the body is very strict to regulation of osteoclastogenesis. Mobilization and differentiation of osteoclast maturation is a complex and sophisticated multi-level regulatory processes. In the relevant regulatory mechanisms, OPG/RANKL/RANK system plays a pivotal role in the process of osteoclast differentiation and maturation. Recent studies revealed that immune cells and osteoclasts were closely connect with each other in the field of bone metabolism, also provide a new therapeutic target for the treatment of bone diseases. The apoptosis of osteoclasts in bone metabolism have been payed more attention,while its mechanism is still not clear, which need further research.
Animals ; Cell Differentiation ; Gene Expression Regulation ; Humans ; Osteoclasts ; cytology ; metabolism ; Osteoprotegerin ; genetics ; metabolism ; RANK Ligand ; metabolism ; Receptor Activator of Nuclear Factor-kappa B ; genetics ; metabolism

OBJECTIVEHuman herpesvirus 6 (HHV-6) isolates are classified into two variants, HHV-6A and HHV-6B, based on distinct genetic, antigenic and biological characteristics. HHV-6 has been associated with encephalitis in children recently. This study aimed to establish a real time PCR assay for simultaneous detection of the two subtypes of HHV-6, and apply this new assay to children with suspected encephalitis, then analyze the relationship between the infection with HHV-6 and encephalitis in children.

METHODThe universal primers and variant-specific TaqMan probes were designed based on the highly conserved sequences of the DNA polymerase gene (U38) of HHV-6. The 5' end of the probes for HHV-6A and HHV-6B was labeled with the fluorescein reporter tetrachloro-6-carboxyfluorescein and 6-carboxyfluorescein (6-FAM), separately, while the 3' end were quenched with 6-carboxy-tetramethylrhodamine. The real time PCR assay for simultaneous detection of HHV-6A and HHV-6B was established. Then, the plasmids of HHV-6A and -6B which were diluted by a 10-fold series from 10(9) to 10(0) copies/microl, together with controls were used for testing both sensitivity and specificity of the real time PCR assay. The cerebrospinal fluid (CSF) specimens from 445 cases of suspected encephalitis were tested with this real time PCR and positive samples were then sequenced.

RESULTBoth HHV-6A (strain ZJ-159) and HHV-6B (strain GS) were positive on the real time PCR assay. There were no cross-reaction with herpes simplex virus type 1, type 2 (HSV-1, HSV-2), varicella-zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), hepatitis B virus, Staphylococcus aureus, Mycoplasma pneumoniae and human DNA. A linear regression curve was obtained when plotting Ct values against the log10 of the viral DNA input for both subtypes of HHV-6. The sensitivity threshold was 10 copies/microl for the real time PCR. HHV-6 positive rate by the real time PCR assay was 4.72% (21/445), including 4 cases with HHV-6A infection, 16 cases of HHV-6B infection and 1 case with mixed HHV-6A and HHV-6B infection. The new PCR assay usually took 2 to 3 hours to provide results.

CONCLUSIONThis new real time PCR assay can simultaneously detect both subtypes of HHV-6, and have high specificity and sensitivity. It will provide an early and sensitive diagnosis of HHV-6 encephalitis in children.

Adolescent ; Child ; Child, Preschool ; DNA Fingerprinting ; DNA Primers ; DNA, Viral ; Encephalitis, Viral ; cerebrospinal fluid ; diagnosis ; virology ; Female ; Fluorometry ; Genotype ; Herpesvirus 6, Human ; genetics ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Alterations of the autophagy-lysosomal pathway (ALP) and autophagy have been involved in lung ischemia-reperfusion (I/R) injury. However, dynamic imaging of ALP function under lung I/R injury particularly is not fully understood. Here we depicted the live-cell fluorescence imaging of autophagosome to monitor ALP activation and autophagy function. The pAsRed2-N1-LC3 vectors were transfected into CRL-2192 NR8383 (an alveolar macrophage cell line) and CCL149 (an alveolar epithelial cell line) successfully. 0-h, 2-h, 4-h, and 6-h hypoxia/0-h, 2-h, 4-h, and 6-h reoxygenation were then induced with an ALP inhibitor (3-MA) or activator (rapamycin) in the culture of transfected cells separately. ALP activation was conformed by up-regulating AMPK and beclin1 expression. Apoptosis was not obvious in 2-h hypoxia/2-h reoxygenation. pAsRed2-N1-LC3 CCL149 and pAsRed2-N1-LC3 NR8383 cells revealed gradually enhanced AsRed2 from 2-h to 6-h hypoxia/reoxygenation. AsRed2 varied sensitively to 3-MA and rapamycin interventions during 2-h hypoxia/reoxygenation. Our data provides a simple method of autophagosome imaging to monitor ALP activation and autophagy function in lung I/R injury.
Animals ; Autophagy ; Base Sequence ; DNA Primers ; Hypoxia ; physiopathology ; In Vitro Techniques ; Lung ; physiopathology ; Lysosomes ; physiology ; Oxygen Inhalation Therapy ; Rats ; Real-Time Polymerase Chain Reaction

OBJECTIVETo study the bacterial community structure of the microbiota in the vaginal fluid from patients with bacterial vaginosis.

METHODSThe composition of bacteria in the samples of vaginal fluid from 3 patients with bacterial vaginosis and 1 normal premenopausal control was investigated by amplified ribosomal DNA restriction analysis(ARDRA).

RESULTSLactobacillus species were the predominant bacteria in the woman without bacterial vaginosis. Bacterial vaginosis was associated with higher concentrations of a variety of bacterial groups. Women with bacterial vaginosis had greater bacterial diversity, with 31 to 37 OTUs operational taxonomic units detected per sample. The species associated with bacterial vaginosis were Leptotrichia, Prevotella sp. and Megasphaera including several species with no close cultivated relatives.

CONCLUSIONSWomen with bacterial vaginosis have complex vaginal infections with many newly recognized species. ARDRA allows rapid analysis of the diversity of microorganisms in the vagina, and is capable of identifying potentially pathogenic bacteria that can not be identified by general culture.

Adult ; Bacteria ; classification ; genetics ; isolation & purification ; Bacterial Typing Techniques ; methods ; DNA, Ribosomal ; analysis ; genetics ; Female ; Humans ; Leptotrichia ; genetics ; isolation & purification ; Megasphaera ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods ; Phylogeny ; Prevotella ; genetics ; isolation & purification ; Restriction Mapping ; Vaginosis, Bacterial ; microbiology

OBJECTIVETo study the genetic diversity of medicinal Dendrobium by SRAP.

METHODThe genetic diversity of 9 spices Dendrobium was studied by using the optimized SRAP reaction system. The NTSYS software was used to analyze the markers.

RESULTForty primer pairs were selected from 88 amplified 1 782 polymorphic bands with an average of 44.55 polymorphic bands per primer pair. Cluster analysis using UPGMA method based on the data of SRAP amplified bands by 40 primer pairs showed that 9 spices of could be distinguished into two main groups. Jaccard's similarity coefficient ranged from 0.330 2-0.789 2.

CONCLUSIONThe results of this research indicate that SRAP molecular marker is efficient to study the medical Dendrobium genetic diversity.

Dendrobium ; classification ; genetics ; Genetic Variation ; genetics ; Nucleic Acid Amplification Techniques ; methods ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction