Comparative epidemiology and virology of fatal and nonfatal cases of hand, foot and mouth disease in mainland China from 2008 to 2014.

This study aimed to analyze the epidemiology and virology of fatal and nonfatal hand, foot, and mouth disease (HFMD) cases in Mainland China. A total of 10,714,237 survivors and 3046 deaths were reported from 2008 to 2014 June, with a case fatality rate of 0.03%. The morbidity of the survivors increased from 37.6/100,000 in 2008 to 139.6/100,000 in 2013 and peaked in 2012 at 166.8/100,000. However, the mortality varied around 0.03-0.04/100,000 across the time. Most of the survivors were distributed in the southern and eastern China, predominantly in the Guangxi and Hainan Province, whereas deaths were dominant in southern (Guangxi) and southwestern (Guizhou) China. The two groups showed similar seasonal fluctuations from 2008 to 2014, peaking in spring and early summer. Of the total cases, 93.97% were children less than 5 years of age, with those ≤ 2 years old accounting for 60.08% versus 84.02% in the survivor and death groups, respectively. Boys were at higher risk of infection than girls in both groups. Five years of virological surveillance showed that 43.73%, 22.04%, and 34.22% of HFMD cases were due to EV71, CoxA16 and other enteroviruses, respectively. EV71 was encountered in most deaths, with no substantial effect of age, gender, month, and year on incidence. Subgenotype C4a was the prevalent EV71 strain in Mainland China, with no significant difference in the VP1 gene related to virulence between the two groups. In conclusion, based on the largest population study, fatal and nonfatal HFMD cases, mainly caused by C4a of EV71, are circulating in Mainland China with a low-cause fatality rate.

[Genetic characteristics of VP1 region of coxsackievirus A10 strains in Ningxia Hui Autonomous Region during 2013-2014].

OBJECTIVE: To investigate the genetic characteristics of coxsackievirus A10(CV-A10) strains isolated from hand, foot and mouth disease (HFMD) cases in Ningxia province. METHODS: Based on the HFMD laboratory network surveillance system, 2 470 patients clinical specimens including 450 faeces and 2 020 throat swaps were collected from various regions people's hospital in Ningxia Hui Autonomous Region during January, 2013 to December, 2014. All specimens were isolated using rhabdomyosarcoma cells. VP1 regional gene of isolated strains was amplified by RT-PCR using degenerate primers and sequenced. Sequences were compared with the database of GenBank by the Blast algorithm to identify the enterovirus genotypes. All the CV-A10 strains were performed the homology and phylogenetic evolution analysis. RESULTS: 450 specimens identified as non-EV-A71, non-CV-A16 enterovirus were collected and 36 CV-A10 strains were isolated, 6 strains were isolated in 2013 and 30 strains were isolated in 2014. The homology of nucleotides and amino acids among 36 CV-A10 strains were 90.6%-100.0% , and 90.2%-100.0%, respectively. Compared 36 strains with genotype A, B, C, D representative strains, it has the highest homology with the genotype C, the nucleotide and amino acids homogeneity were 90.2%-98.9% and 95.7%-99.7%. The phylogenetic tree showed 36 strains and genotype C representative strains located in the same evolutionary branch. CONCLUSION: CV-A10 was one of the most common pathogen of HFMD in Ningxia Hui Autonomous Region. All CV-A10 strains belonged to genotype C and contained wide homology range.

Strong influence of surfactants on virgin hydrophobic microplastics adsorbing ionic organic pollutants.

Microplastic (MP) pollution has become an area of increasing concern because MPs accumulate various types of pollutants. Many previous studies have explored the interactions between MPs and hydrophobic pollutants. However, little research has been conducted on hydrophilic pollutants, which are of much higher concentration and ubiquitous in environment. Surfactants cause hydrophobic MPs to become hydrophilic, which may significantly enhance their capacities to adsorb hydrophilic pollutants. This study explored the influence of co-existing surfactants on the adsorption of ionic organic pollutants by MPs, and found that the presence of an ionic surfactant could significantly enhance the capacity of polyvinyl chloride (PVC, 0.2 mm) MPs to adsorb pollutants with opposite charges. The Langmuir methylene blue adsorption capacity of PVC could be increased from 172 to 4417 ppm in the presence of a sodium dodecyl benzene sulfonate surfactant. Nonionic surfactants impeded the adsorption of both cationic and anionic pollutants due to the steric resistance of the hydrophilic polyethelene glycol chains. The electrostatic interaction mechanism dominated the interfacial behaviors of ionic pollutants on surfactant-adsorbed MP interfaces. The effects of the surfactants were further verified using four different model pollutants and six surfactants. The adsorption capacities of real environmental MPs, including PVC, polyethylene (PE), polypropylene (PP), and polystyrene (PS), increased by three to twenty-six times. The adsorption properties of MPs may be determined by the presence of co-existing surfactants, rather than their polymer species or additives.

Surfactant stealth effect of microplastics in traditional coagulation process observed via 3-D fluorescence imaging.

Microplastics (MPs) have aroused rising social concerns. Although amounts of surfactants exist in wastewater and are expected to alter the surface properties of MPs significantly as they are designed to be adsorbed by hydrophobic particles. However, rare works have been done on the influence of surfactants on the coagulation removal process of MPs which was thought to be an effective way to remove MPs together with other natural particles, such as clay. We used 3-D fluorescence imaging to track the coagulation removal process of polystyrene MPs. Our results indicate that nonionic surfactant, tween 20 in ppm scale, could inhibit the coagulation removal of polystyrene MPs significantly. Residue MPs in the effluent is proportional with the surfactant concentration and increases up to tens of times, which will lead to a dramatic increase in their potential environmental risks. Apparent size effect exists in the coagulation in which smaller MPs can escape from the coagulation removal more easily. Mechanism study suggests that the steric resistance of the hydrophilic flexible polyethylene glycol (PEG) layer formed by tween 20 adsorbed on MP surface inhibits clay deposition and thus hinders subsequent agglomeration and precipitation. A surfactant stealth effect, which is used in the design of nanomedicine to avoid the human immune recognition and clearance of nano-drugs from blood circulation, also exists in the coagulation removal process of MPs. Our finding not only proves the strong influence of surfactants on MPs but also will stimulate related studies on other latent surfactant effects of MPs.

[Preparation and drug releasing property of curcumin nanoparticles].

OBJECTIVE: To prepare curcumin nanoparticles and evaluate the in vitro release of curcumin. METHODS: The chitosan-graft-vinyl acetate copolymers were synthesized by free radical polymerization. Curcumin nanoparticles were synthesized by ultrasonic irradiation. The encapsulation efficiency of the nanoparticles and the in vitro release of curcumin were studied. RESULTS: The nanoparticles were discrete and uniform spheres, covered with positive charges. The encapsulation efficiency of nanoparticles was up to 91.6%. The in vitro release profile showed the slower release rate of curcumin. CONCLUSION: The methods is simple. The nanoparticles possess good physical performance and sustained release character in vitro.

[Genetic Characterization of Coxsackievirus A6 Strains Isolated in Ningxia Region, China, 2013～2015].

To study the genetic characteristics of coxsackievirus (CV)-A6 strains isolated from hand, food and mouth(HFMD)cases in Ningxia Province, China, in 2013~2015.A total of 998 specimens identified as non-EV71,non-CVA16 enteroviruses by real-time polymerase chain reaction were collected in 2013~2015and cultured using RD cells. The viral protein(VP)1gene of isolated strains was amplified by reverse transcription-polymerase chain reaction with degenerate primers and sequenced.Sequences were compared using the GenBank database by the BLAST algorithm to identify virus genotypes. All CV-A6 stains identified underwent homologous comparison and phylogenetic analyses. A total of 227 virus strains were isolated from 998 clinical specimens, and 61 stains were identified as CV-A6.Homologies of nucleotides and amino acids among Ningxia CV-A6 strains were 96.1%~99.8% and 98%~100%,respectively.The nucleotide homogeneity of Ningxia CV-A6 strains with Gudla strains, and the similarity of nucleotides and amino-acid sequences with VP1 of CV-A6 were 82.0%~83.5% and 93.8%~95.7%,respectively.CVA6 was the main pathogen of HFMD apart from EV71 and CV-A16 in Ningxia Province in 2013~2015.

[Genetic characteristics of VP1 region of coxsackievirus A10 strains isolated from hand foot and mouth disease patients in Ningxia Hui Autonomous Region, 2013].

OBJECTIVE: To study the genetic characteristics of VP1 region of coxsackievirus A10 (Cox A10) strains isolated from hand foot and mouth disease (HFMD) cases in Ningxia Hui Autonomous Region (Ningxia) in 2013. METHODS: A total of 280 specimens, which were identified as non-enterovirus 71 and non-Cox A16 by real-time PCR, were collected and cultured by using RD cell, and the VP1 genes of isolated strains were amplified by using reverse transcriptase PCR (RT-PCR) with degenerated primers and sequenced. The sequencing results were aligned with the sequences in GenBank with BLAST algorithm to identify the virus genotypes. Homologous comparison and phylogenetic analysis were conducted for all the Cox A10 strains identified. RESULTS: Among 36 virus strains isolated from 280 clinical specimens, 6 were identified as Cox A10. The homologies of nucleotide and amino acid of the Cox A10 strains isolated in Ningxia were 97.0%-99.8% and 99.0%-99.7% respectively, and the Cox A10 strains isolated in Ningxia shared 76.3%-77.2%, 81.6%-83.1%, 94.4%-98.9% and 80.0%-82.3% nucleotide homologies respectively and shared 92.3%-93.0%, 94.0%-95.3%, 98.0%-99.7% and 90.6%-94.0% amino acid homologies respectively with the representative strains of A, B, C and D genotypes. Phylogenetic tree analysis revealed that Cox A10 strains isolated in Ningxia belonged to genotype C. CONCLUSION: Cox A10 is one of the most common pathogen causing HFMD in Ningxia in 2013. All the Cox A10 stains isolated from HFMD patients in Ningxia belonged to genotype C.

Kindler syndrome protein Kindlin-1 is mainly expressed in adult tissues originating from ectoderm/endoderm.

Mutations of integrin-interacting protein Kindlin-1 cause Kindler syndrome and deregulation of Kindlin-1 is implicated in human cancers. The Kindlin-1-related diseases are confined in limited tissue types. However, Kindlin-1 tissue distribution and the dogma that governs Kindlin-1 expression in normal human body are elusive. This study examined Kindlin-1 expression in normal human adult organs, human and mouse embryonic organs by immunohistochemical analyses. We identified a general principle that the level of Kindlin-1 expression in tissues is tightly correlated with the corresponding germ layers from which these tissues originate. We compared the expression of Kindlin-1 with Kindlin-2 and found that Kindlin-1 is highly expressed in epithelial tissues derived from ectoderm and endoderm, whereas Kindlin-2 is mainly expressed in mesoderm-derived tissues. Likewise, Kindlin-1 was also found highly expressed in endoderm/ectoderm-derived tissues in human and mouse embryos. Our findings indicate that Kindlin-1 may play an importance role in the development of endoderm/ectoderm related tissues.

[Genetic characterization of enterovirus 71 stains isolated in Ningxia province in 2008].

OBJECTIVE: To study the genetic characterization of enterovirus 71 (EV71) strains isolated from specimens of patients with hand-foot-mouth disease (HFMD) in Ningxia province in 2008. METHODS: All the stool, throat swab and vesicle samples that collected from patients with HFMD were cultured. The positive isolates were identified by reverse transcriptase PCR (RT-PCR) with specific primers of EV71. Complete VP1 gene sequences (891 nucleotides) of 29 strains (part of 93 EV71 strains) were determined and compared with A, B and C genotype reference EV71 strains while EV71 China isolates by homogeneity and phylogenetic tree analyses. RESULTS: 215 strains of EV were isolated from 439 specimens. Results from RT-PCR indicated that 93 strains belong to EV71. Phylogenetic tree analysis revealed that the selected 29 stains were clustered with reference strains of C4 subgenotype. The nucleotide identity with C4 reference strains was 91.7%-99.4%. The amino acid homogeneity was 96.6%-100.0%. CONCLUSION: The recently identified EV71 strains in Ningxia province belonged to subgenotype C4 which resembled to most of the isolates in China.

[Anti-tumor effect of monkshood polysaccharide with adriamycin long circulating temperature-sensitive liposome and its mechanism].

AIM: To observe the synergic action of monkshood polysaccharide (MPS) and adriamycin (ADM) long circulating temperature-sensitive liposome (ALTSL) in targeting therapy for H22 tumor-bearing mice and explore the mechanism. METHODS: The anti-tumor activity was evaluated by using the tumor's weight as an index. The life prolongation rate of mice was calculated according to the survival time of the tumor-bearing mice. The killer activity of NK cells and the lymphocyte transformation rate were detected by the LDH release assay and MTT colorimetry, respectively. The apoptosis of tumor cells and the expressions of p53, Fas, Fas-L and caspase-3 were analyzed by flow cytometry. The expressions of IL-2 mRNA and IL-12 mRNA in spleen lymphocytes were determined by RT-PCR. The pathologic changes of tumor, heart, liver and kidney tissues of the tumor-bearing mice were observed under light microscope. RESULTS: Compared with the adriamycin liposome group, the anti-tumor effects were enhanced in (MPS+ALTSL) group with tumor growth inhibitory rate up to 80.4%. The survival time of the tumor-bearing mice in ALTSL and (MPS+ALTSL) groups was significantly prolonged compared with the ADM group (P<0.01). The killer activity of NK cells was higher in ALTSL group than in the NS and ADM groups, and was highest in (MPS+ALTSL) group. The lymphocyte transformation rate of (MPS+ALTSL) group was markedly increased (P<0.01) as compared with the ADM group. The result of RT-PCR indicated that the expressions of IL-2 mRNA and IL-12 mRNA in lymphocytes in the adriamycin long circulating liposome (ALCL) group were significantly higher than those in the ADM group. Expressions of IL-2 mRNA and IL-12 mRNA was much higher in (MPS+ALTSL) group than in ALTSL group. The pathological examination indicated that in (MPS+ALTSL) group, more lymphocytes and monocytes were found in tumor tissue. CONCLUSION: ALTSL can increase the anti-tumor effect and decrease the side-effects (such as the cytotoxicity) of ADM. MPS combined with ALTSL can enhance killer activity of NK cells and transformation of T cells, supporting their synergic anti-tumor effect.

[Synergistic role between rhIL-2 and adriamycin long circulating temperature-sensitive liposome in targeting therapy on tumor].

AIM: To observe the synergistic role between rhIL-2 and adriamycin long circulating temperature-sensitive liposome (ALTSL) in targeting therapy of H22 tumor-bearing mice and explore their anti-tumor mechanism. METHODS: The antitumor activity was evaluated by using the tumor's weight as an index. The prolongation rate of mouse life was calculated according to the survival time of the tumor-bearing mice. The killer activity of NK cells and the lymphocyte transformation rate were detected by the LDH and MTT colorimetry, respectively. The apoptosis of tumor cells and the expression of p53, Fas, Fas-L and Caspase-3 were analyzed by flow cytometry (FCM). The expression of IL-2 mRNA and IL-12 mRNA in splenocytes was determined by RT-PCR. The pathologic changes of tumor, heart, liver and kidney tissues of the tumor-bearing mice were observed under light microscope. RESULTS: The tumoristatic rate of rhIL-2+ALTSL (73.5%) was higher than that of adriamycin liposome (ADML) group (67.0%). The survival time of tumor-bearing mice in ALTSL and rhIL-2+ALTSL groups was significantly extended as compared with the NS group (treated with normal saline) and the free ADM group (P <0.01 or P <0.05). The killer activities of NK cells of ALTSL group and rhIL-2+ALTSL group were higher than those of the NS and free ADM groups, and was highest in rhIL-2+ALTSL group. The lymphocyte transformation rate of ALTSL+rhIL-2 group markedly increased ( P <0.01) as compared with the free ADM group. The result of RT-PCR indicated that the expression of IL-2 mRNA and IL-12 mRNA in splenocytes in the adriamycin long circulating liposome (ALCL) group was significantly higher than that in the free ADM group. The enhancement of rhIL-2+ALTSL on expression of IL-2 mRNA and IL-12 mRNA was much stronger than that of ALTSL alone. The pathological examination indicated that in rhIL-2+ALTSL group, the tumor cells were mostly destroyed, and a large amount of lymphocytes and monocytes were found in tumor tissue. CONCLUSION: ALTSL can increase the anti-tumor effect and decreased the side-effects (such as the cytotoxicity) of ADM. rhIL-2+ALTSL can induce the apoptosis of tumor cells and enhance killer activities of T cells and NK cells. rhIL-2 and ALTSL can synergistically play the antitumor effect.