Adolescent ; Adult ; Bleomycin ; therapeutic use ; Child ; Child, Preschool ; Female ; Hemangioma ; drug therapy ; Humans ; Laryngeal Neoplasms ; drug therapy ; Male ; Middle Aged ; Pharyngeal Neoplasms ; drug therapy ; Young Adult

This study is designed to serve as a reference for the establishment of health security systems for children’s critical diseases. Through analysis of the operation of Shanghai Children Hospital Care Aid (SCHCA), this study explored the financing model and management of a children’s critical disease healthcare system and analyzed the possibility of expanding this system to other areas. It is found that a premium as low as RMB 7 per capita per year under SCHCA can provide high-level security for children’s critical diseases. With the good experience in Shanghai and based on the current basic medical insurance system for urban residents and the new rural cooperative medical scheme (NRCMS), it is necessary and feasible to build a health security system for children’s critical diseases at the national level.
Adolescent ; Child ; Child Welfare ; Child, Preschool ; China ; Delivery of Health Care ; Health Policy ; economics ; legislation & jurisprudence ; Humans ; Infant ; Infant, Newborn

To investigate the production status and the safety influence factors of wolfberry in China. We investigated the detailed factors which affect the quality safe of wolfberry in the periods of July-August 2013 and July-September 2009. The factors include fertilizing patterns, the used pesticide and preliminary process wolfberry. The factors were discussed according to the results of investigation, and suggestions were proposed for the management and production departments of wolfberry.
China ; Fertilizers ; analysis ; Lycium ; chemistry ; growth & development ; microbiology ; parasitology ; Pest Control ; Plant Diseases ; microbiology ; parasitology ; prevention & control

OBJECTIVETo investigate the controlling effects of bionic glue on Paratrioza sinica.

METHODP. sinica and bionic glue were chosen as materials to investigate the adhesive rate, mortality rate, and study the effects of behavior of P. sinica and growth of leaves sprayed with bionic glue.

RESULTSpraying of the bionic glue can significantly increase the adhesive rate of P. sinica with no obviously repellency, and it can be used to control the adults of P sinaca in field with no significant effect on the growth of wolfberry leaves.

CONCLUSIONBionic glue has significant controlling effects on adults of P. sinica, it can be used as an eliminator of adults of P. sinica in field at the beginning of the vegetation season to suppress of the development of P. sinica population.

Adhesives ; pharmacology ; Animals ; Hemiptera ; drug effects ; Insect Control ; Insecticides ; pharmacology ; Lycium ; drug effects ; parasitology ; Plant Diseases ; prevention & control ; Plant Leaves ; drug effects ; parasitology ; ultrastructure

OBJECTIVETo explore the HSVtk gene expression mediated by the retroviral vector and to obtain high titer recombinant retroviral virus.

METHODSThe recombinant vector pRevTRE/HSVtk was constructed by inserting HSVtk gene into pRevTRE. The recombinant retrovirus, which was produced from cloned PA317 cells screened by hygromycin B after "micro-pingpong" technique transferring with pRevTRE/HSVtk plasmids DNA by using modified calcium phosphate precipitation method. HSVtk gene expression was performed on target cells and virus titers were detected in different cultured temper, time and sodium butyrate concentration.

RESULTSThe recombinant retroviral vector pRevTRE/HSVtk was constructed and HSVtk gene expression was detected on target cells after they were infected with the recombinant retrovirus.

CONCLUSIONHigh titer of retroviruses could be obtained in the culture medium of PA317 cell line through "micro-pingpong" technique at 30 hours and 10 mmol/L sodium butyrate concentration followed by frozen ultrafiltration.

Animals ; Breast Neoplasms ; enzymology ; pathology ; virology ; Cell Line ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Enzymologic ; Genetic Vectors ; Humans ; Mice ; NIH 3T3 Cells ; Recombination, Genetic ; Retroviridae ; genetics ; Simplexvirus ; enzymology ; genetics ; Thymidine Kinase ; biosynthesis ; genetics ; Titrimetry ; Transfection

In order to investigate the application of recombinant adeno-associated virus (rAAV) vector containing Tet regulation system and HSVtk gene in cancer gene therapy, pAAV/TRE/HSVtk/Tet-On was constructed and identified with PCR and restriction enzyme digestion. Packaging cells HEK293 were cotransfected with plasmids pAAV/TRE/HSVtk/Tet-On, pAAV-RC and pAAV-helper to produce infectious rAAV, and CsCl2 densitygradient centrifugation method was performed for purification and concentration of rAAV. The viruses were then transduced into MCF-7 cells. The results of dot blot hybridization indicate that the rAAV can transfer the target gene into MCF-7 cells. MTT assay showed that GCV could kill AAV-infected MCF-7 cells under the induction of Dox. The data demonstrated that rAAV containing Tet regulation system and HSVtk gene was successfully obtained, and could be used for further investigation of in vivo and in vitro experiments.
Cell Line, Tumor ; Dependovirus ; genetics ; metabolism ; Doxycycline ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Simplexvirus ; enzymology ; genetics ; Thymidine Kinase ; genetics ; Transfection

BACKGROUNDRevTet-On gene expression system was used to deliver the suicide gene tk to human breast cancer cell line MCF-7 and control the tk gene expression level. The animal model of human breast cancer on severe combined immune deficiency (SCID) mice was set up to explore the suicide gene therapy by the regulation of Tet-On.

METHODSHerpes simplex virus-thymidine kinase (HSVtk) gene was inserted into the plasmid pRevTRE and the recombinant retroviral vector pRevTRE/HSVtk was constructed. Using modified calcium phosphate co-precipitation method, two transfections, pRevTRE/HSVtk and pRevTet-On were performed for MCF-7 cell line and selected by hygromycin B and G418. MCF-7 cell line that stably expressed Tet-regulated tk gene was established. HSVtk gene expression in the MCF/TRE/tk/Tet-On cell line was under the control of Doxycycline (Dox). Cell viability was also determined by MTT assay, whereas HSVtk gene expression was analyzed by reverse transcription-PCR (RT-PCR).

RESULTSMCF/TRE/tk/Tet-On cell survival rate was decreased from 100% to less than 20% when ganciclovir (GCV) concentration was increased from 0 to 1000 microg/ml at 1 microg/ml of Dox after 72 hours of GCV administration. At 1 microg/ml of GCV concentration, the cell numbers decreased from 7 x 10(4) cells/ml to 2 x 10(4) cells/ml when Dox concentration was increased from 0 to 1500 ng/ml after 72 hours culture. In addition, bystander effects were generated in vitro when 10% - 25% of transduced MCF-7 cells were mixed in untransduced MCF-7 cells. On the other hand, the human breast cancer models in SCID mice were set up. The tk gene was expressed with the regulated character after MCF/TRE/tk/Tet-On cells were implanted into the female SCID mice 7 days after Dox induction followed by intraperitoneally administration of GCV for 23 days. Subcutaneous tumors in SCID mice that were implanted with MCF/TRE/tk/Tet-On cells shrank remarkably after Dox and GCV administration as compared with the control.

CONCLUSIONThe human breast tumor cells (MCF-7) expressing HSVtk gene can be eradicated by administration of GCV and induced with tetracycline or its derivative Dox in vitro and in vivo.

Animals ; Breast Neoplasms ; therapy ; Bystander Effect ; Cell Line, Tumor ; Cell Survival ; Doxycycline ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; Genetic Therapy ; methods ; Genetic Vectors ; Herpesviridae ; genetics ; Humans ; Mice ; Mice, SCID ; Retroviridae ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Thymidine Kinase ; genetics ; Transfection

Objective To evaluate the association between aortic arterial stiffness and target organ damage, including renal function decline and left ventricular diastolic dysfunction among elderly diabetic patients and to provide theoretical basis for early prevention and intervention. Methods Brachial-ankle-artery pulse wave velocity(baPWV)reflecting aortic arterial stiffness was observed in 151 diabetic patients with(n=78)or without hypertension(n=73). Clinical data were collected, cardiac structure and function were assessed by echocardiography. Estimated glomerular filtration rate(eGFR)was calculated by CKD-EPI fomula. Results Of 151 subjects, the average age was(75.05±10.13)years and the average baPWV values was (1 803.85±491.04)mm/s. Patients with both diabetes and hypertension had significantly higher baPWV values[(2006.49± 536.70)mm/s]than that of diabetic patients[(1596.96 ±331.02)mm/s, P<0.01]. Multiple linear regression analysis demonstrated that baPWV was negatively associated with E/A ratio and eGFR(β'=-0.249, -0.225, P<0.05), but positively associated with hypertension(β'=2.008, P<0.05). By applying area under the receiver operator characteristic curve, baPWV had powerful predictive power for renal impairment(AUC=0.695, 95% CI: 0.548-0.842)and left ventricular diastolic dysfunction(AUC=0.687, 95%CI: 0.532-0.842). Conclusion Patients with both diabetes and hypertension had significantly higher baPWV values than that of diabetic patients. Higher baPWV was independently associated with greater burden of sub-clinical disease in renal impairment and left ventricular diastolic dysfunction.

Objective To investigate whether mild hypothermia can promote neurogenesis in the dentate gyrus of hippocampus and cognitive function recovery after traumatic brain injury ( TBI) through inhibiting apoptosis of hippocampal neurons. Methods A total of 66 healthy adult Sprague-Dawley rats were randomly divided into sham group, TBI group and TBI+hypothermia group, with 22 rats in each group. The rat TBI model was established using the fluid percussion device. The rats in TBI +hypothermia group received 4-hour hypothermia therapy immediately after injury, with the target temperature of 33. 5℃. Bromodeoxyuridine (BrdU) was injected into the rats' abdominal cavity to label the mitotic cells. The test of Morris water maze was used to evaluate the rats' spatial learning and memory capabilities. Immunofluorescence staining was used to observe the expression levels of BrdU, doublecortin (DCX), neuron specific nuclear protein (NeuN), cysteinyl aspartate specific proteinase 3 (caspase-3) and cleaved caspase-3 expressions in dentate gyrus of hippocampus at 7 days and 28 days after injury. Expressions apoptosis-related proteins including the factor associated suicide ( FAS )/factor associated suicide ligand (FASL), B-cell lymphoma-2 (Bcl-2), caspase-3 and cleaved caspase-3 expressions were detected by Western blot assay. Results The water maze tests at 28 days after injury showed that compared with TBI group, the escape latency in TBI+hypothermia group was significantly shorter [(24. 2 ± 5. 9)s:(18 ± 4. 1)s], and both the time in the target quadrant and the number of platform crossing were increasedsignificantly[(24.9±6.5)s:(31.7±5.2)s; (1.9±0.8) times:(3.5±1.2)times](P<0. 05). Compared with the sham group, in TBI group and TBI+hypothermia group, the BrdU+ new-born cells in the dentate gyrus of hippocampus were significantly increased at 7 days after injury [(9. 4 ± 4. 1):(33. 4 ± 3. 8);(9. 4 ± 4. 1):(45. 8 ± 5. 6)], the BrdU+ /DCX+ new-born neurons were increased at 7 days after injury [(2. 0 ± 0. 6):(9. 6 ± 1. 6);(2. 0 ± 0. 6):(19. 2 ± 3. 7)], and the BrdU+ /NeuN+mature neurons were increased at 28 days after injury [(2. 6 ± 1. 0) :(17. 2 ± 3. 9); (2. 6 ± 1. 0) :(33. 6 ± 9. 1)] (P<0. 01). TBI group showed more obvious increase than the TBI+hypothermia group (P<0. 01). Moreover, compared with 7 days after injury, the number of BrdU+ cells at 28 days after injury was further increased in TBI +hypothermia group but decreased in TBI group [(45. 8 ± 5. 6) :(58. 8 ± 9. 2);(33. 4 ± 3. 8):(22. 0 ± 3. 5)](P<0. 05 or <0. 01). Compared with the sham group, the caspase-3 +NeuN+ and caspase-3 +NeuN+ apoptotic neurons were significantly increased at 7 days after injury in TBI group [(2. 0 ± 0. 9):(11. 6 ± 2. 6); (2. 6 ± 1. 0):(10. 2 ± 2. 9)] (P<0. 05). Compared with the TBI group, the cleaved caspase-3 +NeuN+ apoptotic neurons were decreased in TBI+hypothermia group [(6. 6 ± 2. 0):(11. 6 ± 2. 6)](P<0. 05). Furthermore, compared with the TBI group, mild hypothermia might down-regulate the expression of FAS, FASL, cleaved caspase-3 and caspase-3 and up-regulate the expression of Bcl-2 in the hippocampus [(1. 54 ± 0. 15) :(1. 14 ± 0. 12);(1. 06 ± 0. 04):(0. 80 ± 0. 09); (0. 84 ± 0. 03):(0. 62 ± 0. 08); (0. 93 ± 0. 06):(0. 86 ± 0. 09);(0. 71 ± 0. 01):(1. 58 ± 0. 18)](P<0. 05). Conclusions Mild hypothermia might inhibit apoptosis of hippocampal neurons through cleaved caspase-3, FAS/FASL and Bcl-2 pathways, thus improving the neurogenesis and maturation of neurons in the dentate gyrus of hippocampus and facilitating cognitive function recovery in rats. It indicates that the function of hypothermia in anti-apoptosis and neurogenesis and maturity of hippocampal neurons may have a potential role in predicting the prognosis of TBI patients.

Objective To analyze genetic characteristics of hemagglutinin( HA) gene of H9N2 avian influenza viruses( AIVs) circulating in Anhui Province from 2013 to 2018． Methods All H9N2 positive samples tested by real－time polymerase chain reaction( PCＲ) were inoculated into specific patho- gen free ( SPF) chicken eggs for isolation and purification． Viral ＲNA was reversely transcribed into cD- NA and then amplified with gene specific primers． PCＲ products were sequenced and the gene sequences were analyzed using molecular and bioinformatics software． The DATAMONKEY online server was conducted to analyze selection pressure，and protein structure homology modelling was computed by the SWISS－MODEL server． Ｒesults 33 H9N2 AIVs isolated from live poultry markets belonged to h9．4．2．5 in the phylogenetic tree． The receptor binding sites of HA gene at 183，226 and 227 position were mutated into N，L and M，respectively． Meanwhile 189 and 190 sites presented with genetic polymorphism． Since 2015，all H9N2 viruses in this study carried 6 potential N－linked glycosylation sites． It was found that po- sition 160 of HA gene was subjected greater positive selection pressure，presented 7 spatial conformations at least． Conclusions The H9N2 viruses isolated from live poultry markets in Anhui Province possess the molecular characteristics of infecting mammals and the ability of antigenic drift，so we need to pay more attention to the genetic characteristics of the viruses．