Objective To investigate the effects of acute oxidative stress induced by H2O2 on expression of senescence marker protein30 (SMP30) and morphology,survival rate of human lens epithelial cells (HLECs).Methods HLECs were treated with H2O2(0 μmol · L-1,100 μmol · L-1,200 μmol · L-1,300 μmol · L-1) for 24 hours,the acute oxidative stress models were established,the changes of cell morphology was observed,and MTT was used to analyze the cells state,the expressions of SMP30 were measured by Western blot.Results The cell density decreased,morphological changed and viability of cells significant decreased in 100 μmol · L-1 and 200 μmol ·L-1 treated group,the large and round cells appeared,the cell body stretched with unclear boundary.With the H2O2 concentration increased,the viability of cells were gradually decreased in treated group,there were statistical differences compared with 0 μmol · L-1 treated group (all P ＜ 0.05).The relative expression of SMP30 in control group and 100 μmol · L-1 and 200 μmol · L-1 treated group were 0.273 ±0.055,0.464 ± 0.058,0.442 ± 0.050,respectively.There were significant differences between 100 μmol · L-1,200 μmol · L-1 treated group and control group (all P ＜ 0.05),and there was no statistical difference between 100 μmol · L-1 and 200 μmol · L-1 treated group (P ＞ 0.05).Conclusion SMP30 is up-regulated in HLECs under acute oxidative stress induced by H2O2,the cell morphology is changed,the viability of cells is decreased,and SMP30 may be involved in the process of acute oxidative stress in HLECs.

The prevalence of acute flaccid paralysis (AFP) associated with EV71 and the genetic variation in Fujian , China from 2003 to 2012 was investigated in this study .Descriptive epidemiology was used to analyze the epidemiologic and clinical features of AFP cases associated with EV 71 .Phylogenetic analysis was performed to explore the genetical characteris-tics of EV71 based on the complete VP1 nucleotide and amino acid sequences .Results showed that the mean incidence of EV71-associated AFP in children under 15 years old was 2 .24/10 000 000 in Fujian Province during 2003 and 2012 ,based on the number of EV71 isolates and the reported AFP cases .And the incidence has increased since 2008 .The EV71 strains isolated from the AFP cases or from the healthy contacts were distributed in 9 prefectures of Fujian Province ,most in the months of May and June .Of 76 .0% (19/25) of AFP cases associated with EV 71 were the children under 3 years and the male-to-female ratio was 1 .5 :1 .Twenty out of twenty-two cases (90 .91% ) had fevers before the onset of paralysis .Most cases had unilater-al limb paralysis (14/22 ,63 .6% ) .Typical manifestations of hand-foot-and-mouth disease (HFMD) were observed in five cases before the onset of paralysis .Residual paralysis was observed in two cases during the follow-up visits .The strains isolated from 25 cases belonged to genotype C4 .All other strains belonged to subtype C4a except the subtype C4b strains isolated in 2003 .The homology among the strains was high in 2009-2011 ,and the homology among these strains and the representative strains in Fuyang ,Anhui Province was also in the high level .Therefore ,it was possible that the isolated strains had the same origin and might cause the epidemic .In conclusion ,an AFP surveillance system could be developed for analyzing the incidence of AFP associated with EV71 ,determining the features of the isolates ,and describing the intensity and trends of EV71 epidem-ics .

Objective: To investigate the effect of mitoxantrone (MIT) on calreticulin (CRT) expression in B16 cells, and to observe the immune effect of B16-membrane antigen vaccine highly expressing CRT on B16 tumor-bearing mice. Methods: The expression of CRT on membrane of B16 cells was detected by immunofluorescence after treatment with different concentrations of MIT. B16-implanted mouse model was established, and the growth of B16-implanted tumors and CRT expression in B16-implanted tumor tissues were observed after treatment with different concentrations of MIT. Membrane antigen vaccines from both normal B16 cells and MIT-treated B16 cells were prepared, and mice were immunized before B16 cell implantation. The infiltration of immune cells into B16 tumor tissues and the ratios of CD4~+ and CD8~+ T cells in the spleen of B16 tumor-bearing mice were examined by immunohistochemistry and flow cytometry, respectively. Results: Flow cytometry results showed that MIT dose-dependently increased CRT expression on B16 cell membrane, with CRT expression in control and high dosage MIT groups being (29.40±3.57)% and (72.20±2.94)% (P<0.05), respectively. MIT also increased CRT expression in B16 tumor tissues, with those in the control and high dosage MIT groups being 3.21±1.37 and 9.17±1.06 (P<0.05), respectively. MIT effectively inhibited the growth of B16 tumors (P<0.05). Compared with normal B16 cell membrane antigen vaccine, the vaccine highly expressing CRT increased the numbers of DCs and T cells in B16 tumors tissues and the ratios of CD4~+ and CD8(+) T cells in the spleen (P<0.05). Conclusion: MIT can increase CRT expression on membrane of B16 cells. B16-membrane antigen vaccine highly expressing CRT can enhance the infiltration of DCs and T cells in melanoma, thus improving the immune effect of B16-membrane antigen vaccine.

The Assistive Devices Program was funded and supported sufficiently, and improved the qulity of life of the disabled persons significantly (scores of SF-36). Most disabled users were satisfied with the Program. Some problems, such as inefficient way of working,undefined screening standards, lack of integrity of the assessment content, limited categories of assistive devices and home modifications,unavailable follow-up services, needed to be improved.

Objective To analyze the genetic characteristics of VP1-VP4 genes carried by cox-sackievirus A6 (CVA6) strains isolated from severe cases of hand, foot, and mouth disease (HFMD) in Shenzhen during 2012 to 2015. -ethods The VP1-VP4 genes of CVA6 strains isolated from severe HFMD cases in Shenzhen during 2012 to 2015 were amplified and sequenced. Phylogenetic analysis was performed to analyze the VP1-VP4 genes of CVA6 isolates and sequences downloaded from GenBank by using DNASTAR6. 0 and MEGA6. 02 software packages. Results Four cases of severe HFMD were caused by CVA6 in Shenzhen during 2012 to 2015. All of the patients had the symptom of fever, skin rash and aseptic encephalitis. The CVA6 strain causing severe HFMD in 2013 shared 98. 8％-98. 9％ homology in nucleotide sequences and 99. 3％-99. 8％ in amino acid sequences with the strains isolated in 2012. Two amino acid mutations were found in the CVA6 strain isolated in 2013, which were G73E in VP2 region and S13G in VP1 region. However, the CVA6 strain isolated in 2015 only shared 95. 0％ homology in nucleotide sequences and 99. 3％ homology in amino acid sequences with the strain isolated in 2013. Six amino acid mutations were identified including E73G in VP2 region and T5A, S27N, A30V, N137S and V242I in VP1 region. The phylogenetic analysis revealed that the four CVA6 strains belong to D3 sub-genotype. The CVA6 strains causing severe cases in 2012 had the nearest genetic relationship with the strain isolated in Changsha in 2012 (KJ156349). The CVA6 strain isolated in Shenzhen in 2013 had the nearest genetic relationship with the strain isolated in Shanghai in 2013 (KJ612513). The Shenzhen CVA6 isolate in 2015 showed high similarity to Weifang CVA6 isolate in 2014 (KX752785). Conclusions All CVA6 strains causing severe HFMD ca-ses in Shenzhen during 2012 to 2015 belongs to D3 sub-genotype. Mutations of S27N and A30V in the VP1 region of the CVA6 isolate in 2015 are located in the B cell epitopes. In addition, the VP1-V242I mutation in the CVA6 strain isolated in 2015 is located in the binding site of PSGL-1 receptor. These mutations may affect the binding of CVA6 strains to the cellular receptors and their infectivity to people.

Objective: To investigate the imaging characteristics and the cause for the misdiagnosis and mistreatment of epidermoid cyst in intrapancreatic accessory spleen (ECIPAS) in order to improve the accuracy of preoperative diagnosis. Methods: The clinical and imaging data of 8 patients with ECIPAS confirmed by pathology in Zhejiang Provincial People′s Hospital between June 2008 and February 2018 were collected. The reason for doctor visit included CA19-9 elevation (n=1) and pancreatic occupying mass (n=9) during physical examination and no obvious symptoms were reported. CT and MRI imaging features, diagnosis and treatment were analyzed. Results: The lesions in 8 cases of ECIPAS were all located in the tail of the pancreas and were cystic and solid. The lesions in 3 cases were mainly cystic and the cystic wall was linear, whose CT density, MRI signal and enhancement pattern cannot be compared with those of the spleen. Solid components can be seen in 5 cases, and the CT density or MR signal of the solid part was similar to that of the spleen. After enhancement, the solid part at the artery stage was uniformly enhanced and the enhancement degree was higher than that of the pancreas. Similar to the spleen, it was uniformly enhanced at the portal vein stage and the enhancement degree of the spleen was consistent. All 8 patients were diagnosed with pancreatic neoplastic lesions before surgery, and 1 patient had pancreatic fistula and peripancreatic necrosis after surgery. Postoperative pathology confirmed the diagnosis of ECIPAS. Conclusions Improving the radiologists and clinicians′ cognition of the imaging manifestations of ECIPAS can improve the accuracy of preoperative diagnosis and avoid unnecessary surgery due to misdiagnosis.

Objectiveto investigate the differential expression of intestinal flora and serum metabolites and potential biomarkers in patients with pre-diabetic sputum syndrome. MethodA total of 34 patients with pre-diabetic sputum syndrome were included as the phlegm syndrome group，and 37 healthy people were selected as the normal group. Serum and fecal samples of the two groups were collected，and liquid chromatography-mass spectrometry （LC-MS） non-targeted metabolomics and 16S rRNA high-throughput sequencing technology were used to detect serum metabolites and different intestinal flora of the two groups and explore the relationship among pre-diabetic sputum syndrome，serum metabolites，and intestinal flora. ResultIn the distribution of disease syndrome elements in the phlegm syndrome group，the first five disease syndrome elements in terms of frequency and proportion were dampness （73.53%），Qi stagnation （58.82%），Yin deficiency （50.00%），blood stasis （41.18%），and heat （35.29%）. According to the frequency and proportion of disease location syndrome elements，the first three main disease location syndrome elements were spleen （100.00%），liver （41.18%），and kidney （23.53%）. The results of 16S rRNA high-throughput sequencing showed that there were 44 different intestinal flora between the two groups. In order genus，there were significant differences in Bifidobacterium，Veillonococcus，and Roseococcus between the two groups （P<0.05）. The diversity，abundance，and evenness of intestinal flora in the phlegm syndrome group were lower than those in the normal group，with the difference not statistically significant. There was no significant difference in the community structure between two groups. The results of serum metabolomics showed that there were 13 differential metabolites in the two groups，which were mainly concentrated in amino acid metabolism，bile secretion，bile acid biosynthesis，and lipid metabolism （P<0.05）. The correlation among differential metabolites，intestinal flora，and syndrome elements was analyzed，and the results showed that ① lysine was positively correlated with spleen，Yin deficiency，and blood stasis，while taurocholic acid was positively correlated with liver，kidney，blood stasis，and dampness，and there was a positive correlation between taurocholic acid and yin deficiency and heat. The taurochenodeoxycholic acid was positively correlated with liver and dampness，and there was a negative correlation between arachidonic acid and dampness，as well as a positive correlation between glucose and spleen and blood stasis. ② Clostridium was positively correlated with spleen，kidney，Yin deficiency，and Qi stagnation. Rosepiella was negatively correlated with spleen，and Sutterella was negatively correlated with dampness. Bacteroides was negatively correlated with the spleen and kidney，and Bifidobacterium was negatively correlated with the spleen and dampness. ③ Bifidobacterium was positively correlated with glycine，threonine，lysine，and deoxycholic acid significantly，negatively correlated with cholic acid significantly，and positively correlated with taurochenodeoxycholic acid and pyruvic acid. Clostridium was positively correlated with glycine significantly and positively correlated with threonine and lysine. Lachnospira was negatively correlated with glycine，threonine，and pyruvic acid. Lysine was also negatively correlated with Faecalibacterium and Eubacterium ventriosum and positively correlated with Megamonas. There was a positive correlation between taurocholic acid and glycine bile acid and Campylobacter，between taurochenodeoxycholic acid and Veillonococcus，and between glucose and Rosepiella and Eubacterium ventriosum. There was a negative correlation between pyruvic acid and Escheria-Shigella and between taurochenodeoxycholic acid and Prevotella. Conclusionthere are differences in intestinal flora and serum metabolites between patients with pre-diabetic sputum syndrome and healthy people. The intestinal flora and metabolites have been disturbed in the stage of pre-diabetes，Bifidobacterium，Clostridium，Lachnospira，glycine，threonine，and lysine may be the breakthrough to explore the development of pre-diabetic sputum syndrome.

Interleukin-27 (IL-27), a heterodimeric cytokine, plays a protective role in diabetes. Ghrelin, a gastric hormone, provides a hunger signal to the central nervous system to stimulate food intake. The relationship between IL-27 and ghrelin is still unexplored. Here we investigated that signal transducer and activator of transcription 3 (STAT3)-mechanistic target of rapamycin (mTOR) signaling mediates the suppression of ghrelin induced by IL-27. Co-localization of interleukin 27 receptor subunit alpha (WSX-1) and ghrelin was observed in mouse and human gastric mucosa. Intracerebroventricular injection of IL-27 markedly suppressed ghrelin synthesis and secretion while stimulating STAT3-mTOR signaling in both C57BL/6J mice and high-fat diet-induced-obese mice. IL-27 inhibited the production of ghrelin in mHypoE-N42 cells. Inhibition of mTOR activity induced by siRNA or rapamycin blocked the suppression of ghrelin production induced by IL-27 in mHypoE-N42 cells. siRNA also abolished the inhibitory effect of IL-27 on ghrelin. IL-27 increased the interaction between STAT3 and mTOR in mHypoE-N42 cells. In conclusion, IL-27 suppresses ghrelin production through the STAT3-mTOR dependent mechanism.

The Ly-6 and uPAR (LU) domain-containing proteins represent a large family of cell-surface markers. In particular, mouse Ly-6A/Sca-1 is a widely used marker for various stem cells; however, its human ortholog is missing. In this study, based on a systematic survey and comparative genomic study of mouse and human LU domain-containing proteins, we identified a previously unannotated human gene encoding the candidate ortholog of mouse Ly-6A/Sca-1. This gene, hereby named LY6A, reversely overlaps with a lncRNA gene in the majority of exonic sequences. We found that LY6A is aberrantly expressed in pituitary tumors, but not in normal pituitary tissues, and may contribute to tumorigenesis. Similar to mouse Ly-6A/Sca-1, human LY6A is also upregulated by interferon, suggesting a conserved transcriptional regulatory mechanism between humans and mice. We cloned the full-length LY6A cDNA, whose encoded protein sequence, domain architecture, and exon-intron structures are all well conserved with mouse Ly-6A/Sca-1. Ectopic expression of the LY6A protein in cells demonstrates that it acts the same as mouse Ly-6A/Sca-1 in their processing and glycosylphosphatidylinositol anchoring to the cell membrane. Collectively, these studies unveil a novel human gene encoding a candidate biomarker and provide an interesting model gene for studying gene regulatory and evolutionary mechanisms.
Humans ; Membrane Proteins/genetics* ; Pituitary Neoplasms/genetics* ; Biomarkers