Objective To investigate the effect of high mobility group box-1 protein (HMGB1) on immune function of peritoneal macrophages in mice. Methods The peritoneal macrophages were isolated from female BALBc mice in vitro. The macrophages were stimulated with different doses of HMGB1 (1, 10, 100, 1 000ng/ml) for 6 hours, or with 100ng/ml HMGB1 for different duration (0, 6, 12, 24, 48, 72 hours). The phagocytosis ability of macrophages was determined by the neutral red dye uptake assay and optical densities of samples were read at 540nm. The cytotoxicity to L1210 cells was measured with MTT assay and was expressed by absorbance at 570nm. The chemotaxis was assayed by using the Costar Transwell cell culture chamber inserts and the cells on the lower surface of the transwell membrane were fixed and counted. The cells were harvested at the indicated time points, fixed, and stained for indirect immunofluorescence, then the expressions of I-Ak MHC class Ⅱ alloantigen on macrophages were determined with flow cytometry. Results The effects of HMGB1 on phagocytic activities, cytotoxicity, chemotaxis and I-Ak antigen expression of macrophages were enhanced in a dose-dependent manner, starting at doses as low as 1ng/ml and with a maximal response at 100ng/ml (P

Objective To investigate the relationship between C-reactive protein and clinical significance of Tokyo Guidelines for acute cholangitis.Methods Analyzed 739 cases with acute cholecystitis in Xianyang Central Hospital of Shaanxi province.Records of patients diagnosed with acute cholecystitis were screened between September 2013 and October 2015.The criteria of Tokyo guidelines were used in grading the severity of acute cholecystitis.Patients were divided into 3 groups,Group Ⅰ (n =450),Group Ⅱ (n =260) and Group Ⅲ (n =29).C-reactive protein values at the time of admission were analyzed and compared among the groups.Results Mean C-reactive protein levels of groups were found to be significantly different,18.74 mg/L in Group Ⅰ,133.67 mg/L in Group Ⅱ,and 257.43 mg/L in Group Ⅲ.C-reactive protein values among the groups were found to be highly and significantly correlated with the disease grade (P ＜ 0.01).After evaluating C-reactive protein levels according to the grade of the disease,Group Ⅱ was distinguished from Group Ⅰ with a cutoff C-reactive protein level of 72.54 mg/L,and from Group Ⅲ with a value of 195.85 mg/L.Those results were found to be statistically significant (P ＜ 0.01).Conclusion C-reactive protein can be accepted as a strong predictor in classifying different grades of the disease,and treatment can be reliably planned according to this classification.

AlM: To investigate the efficacy of photodynamic therapy ( PDT) and intravitreal injection with ranibizumab on macular choroidal neovascularization ( CNV ) of pathologic myopia ( PM) .
METHODS: There were patients ( 32 eyes ) who were diagnosed as PM with CNV. Randomly selected 16 cases ( 16 eyes ) which were given the PDT treatment ( PDT group ) . The remaining were given both PDT and intravitreal injection with ranibizumab ( combination group) . There is no significant difference on macular edema between two groups. We analyzed the changes in the best corrected visual activity ( BCVA) , optic coherence tomograph ( OCT ) and fundus fluorescein angiography (FFA) before and 1, 6mo after treatment.
RESULTS:One month after the treatment in PDT group:the BCVA increased while the CMT decreased compared with that of pretreatment (P<0. 05). One month after the treatment in combination group: the BCVA increased while the CMT decreased significantly compared with that of pretreatment (P<0. 01);the changes of BCVA and CMT showed statistically significant between the two groups ( P<0. 05). Six month after the treatment in PDT group: the BCVA increased while the CMT decreased compared with that of pretreatment ( P <0. 05 ). Six month after the treatment in combination group: the BCVA increased while the CMT decreased significantly compared with that of pretreatment ( P <0. 01 ); compared with changes of BCVA and CMT in two groups, the difference was significant after treatment (P<0. 05). Compared 1mo with 6mo after treatment:there was no significant difference in the BCVA and CMT changes (P>0. 05). One month after treatment: in PDT group, FFA showed no leakage or reduced leakage of CNV in 11 eyes (69%), and the fundus remained leaky in 5 eyes ( 31%); in combination group, FFA showed no leakage or reduced leakage of CNV in 13 eyes (81%);the fundus remained leaky in 3 eyes (19%). Six month after treatment:in PDT group, FFA showed no leakage or reduced leakage in 10 eyes ( 62. 5%); the fundus remained leaky in 4 eyes ( 25%); two eyes ( 12. 5%) relapsed leakage; in combination group, FFA showed no leakage or reduced leakage of CNV in 15 eyes (94%);the fundus remained leaky in 1 eye (6%).
CONCLUSlON: Not only PDT but also PDT and intravitreal injection with ranibizumab can block CNV of pathologic myopia completely or partly, and reduce the danger causing descent of vision. Effects and the stability of the combination therapy is superior to PDT treatment.

Objective To obtain the Hiwi gene encoding full-length and construct human Hiwi adenoviral vectors carrying green luorescence protein (GFP), and to establish foundation for a further study on Hiwi function and mechanism of inducing leukemia stem cell differentiation and apoptosis.Methods All coding areas of human Hiwi gene full length were amplified using method of overlapping extension PCR technology, and the full length coding aeras were inserted into the vector of Flag-IRES-hrGFP carrying GFP with Gateway technology to construct pDown-Hiwi-3 × flag-IRES-hrGFP. The cloning vector pDown-Hiwi-3 × flag-IRES-hrGFP and expression vector pAV. Des1d were used for homologous recombination reaction to obtain recombinant adenovirus vector pAV.Ex1d-Hiwi-3× flag-IRES-hrGFP.The positive clones were selected by PCR to extract the recombinant adenovirus plasmid and to pack into recombinant Ad-Hiwi-3 × flag-IRES-hrGFP adenovirus. Results The human recombinant Hiwi was successfully cloned and the recombinant adenovirus vector pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP was found to be successfully constructed via restriction enzyme digestion and sequencing methods. The adenovirus vector pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP was transfected into the HEK293A cells using lipofectamine mediated gene transfection method. Under fluorescence microscope, the transfected cells with green fluorescence could be observed.Conclusion The expression plasmid of adenovirus vector pAV.Ex1d-Hiwi-3 × flag-IRES-hrGFP is successfully constructed and it can express in HEK293A cells.

OBJECTIVE:To study the effect of polysaccharides of Radix scrophulariae on immune function in mice under nor-mal physiological and hypoimmunical state. METHODS:80 mice were randomly divided into normal group,model group,treat-ment high-dose,medium-dose,low-dose groups and physiological high-dose,medium-dose,low-dose groups,10 in each group. Mice in normal group and model group were received distilled water (10 mL/kg) intragastrically,treatment high-dose,medi-um-dose,low-dose groups and physiological high-dose,medium-dose,low-dose groups were administrated 0.16,0.08,0.04 g/kg drug,ig,once a day,for 7 d. From the third day,mice in model group and treatment high-dose,medium-dose,low-dose groups were respectively received cyclophosphamide(100 mg/kg),ip,for 4 d to induce hypoimmunical model. Carbon clearance test was adopted to determine the carbon clearance indexes and organs(chest,spleen)indexes of mice. Another 80 mice were grouped with the same administration,serum half hemolytic value (HC50) was determined after compressed Mianyang red blood cell sensitiza-tion. Then another 80 mice were grouped with the same administration,1% dinitrofluorobenzene method was used to induce de-layed hypersensitivity in mice;its ear swelling was determined,as well as IL-2,IL-4,immunoglobulin G(IgG),immunoglobulin M(IgM),γ-interferon(IFN-γ)contents in serum,and the proliferation in vitro of splenic lymphocytes were detected. RESULTS:High-dose polysaccharides can promote the increasing of immune organ and carbon clearance indexes(P<0.05);enhance the inten-sity of delayed type hypersensitivity (HC50 increasing)(P<0.01) and improve the decreasing of serum hemolysin in model mice (P<0.01);promote the splenic lymphocytes proliferation and increase IL-2,IL-4,IgG,IgM,IFN-γ contents in serum(P<0.05 or P<0.01). CONCLUSIONS:Polysaccharides can enhance the immune function in mice under normal physiological and hypoim-munical state induced by cyclophosphamide.

Objective To identify novel mutations in the GATA6 gene associated with congenital atrial septal defects (ASD).Methods This was a case-control study.A cohort of 220 unrelated Han-race patients with congenital ASD and 200 unrelated ethnically matched healthy individuals used as controls,who were admitted to Tongji University Affiliated Tongji Hospital from January,2007 to October,2011,were recruited.The peripheral venous blood samples from the participants were prepared.All the coding exons and their flanking sequences of the GATA6 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination technique.The acquired sequences were aligned with the sequences derived from GenBank by BLAST to identify the sequence variations.The software ClustalW was used to analyze the conservation of the altered amino acids.Results Three novel heterozygous missense GATA6 mutations,c.250G ＞A (p.A84T),c.649G ＞C (p.G217R) and c.1270A ＞C (p.S424R),were identified in 3 of 220 ASD patients,respectively.None of the three mutations was detected in 200 healthy control individuals.A cross-species alignment of GATA6 encoded protein sequences showed that the mutated amino acids were relatively conserved evolutionarily.Conclusion The identification of novel GATA6 mutations associated with ASD contributes to the reveal of the mechanism involved in the pathogenesis of ASD.

The Qinghai⁃Tibet Plateau is known as one of the highest endemic areas of echinococcosis However，the dog infection rates of Echinococcus granulosus in the plateau regions were similar to other non⁃Tibetan areas with the high endemic，and most of the rates were below 40%. The infected dogs with E. multilocularis were ubiquitous in Ganzi Prefecture of Sichuan Province and Qinghai Province where many survey data were done and available，which was much different from those in non⁃Ti⁃betan areas where the geographical distribution of dogs infected with E. multilocularis was sporadic. The total infection rates of Echinococcus in dogs kept stable from 1983 to 2009 in Ganzi Prefecture of Sichuan Province and did not show much variation from 2000 to 2014 in Qinghai Province as well. Since 2006，the national comprehensive prevention and control strategy and mea⁃ sures against echinococcosis have been launched in China，and significant progress has been made. In the endemic Tibetan area of Sichuan，the general Echinococcus infection rates in dogs were 28.10%，15.87%，19.22%，3.28% and 1.11% from 2009 to 2013，respectively，and the Echinococcus coproantigen⁃positive rate in Gannan Prefecture of Gansu Province and parts of Qing⁃ hai Province also decreased. This paper reviews the literature on the characteristics of dog infections in the Qinghai⁃Tibetan Plateau，so as to provide useful information to support echinococcosis control and prevention there.

Objective Intranuclear forkhead/winged helix transcription factor p3(Foxp3) plays a key role in T cell-mediated immunosuppression.The present study was performed to investigate the effects of high mobility group box-1 protein(HMGB1) on Foxp3 gene as well as protein expressions in splenic regulatory T cells(Tregs) and their potential regulating mechanisms in mice.Methods CD4+CD25+Tregs isolated from the spleens of male BABL/c mice by magnetic beads were seeded on 96-well(1?105 cells/well) cell culture plates coated with anti-CD3(1 ?g/ml) and soluble anti-CD28(1 ?g/ml),and the cells were stimulated with HMGB1 at various intervals or at different concentrations.After being stimulated,the Foxp3 mRNA/protein expressions in the Tregs were determined.The time-dependent and dose-dependent responses between HMGB1 and intranuclear Foxp3 expression were analyzed by flow cytometry,and the expressions of Foxp3 mRNA of Tregs were analyzed by quantitative PCR of SYBR GREEN.Results After stimulation with HMGB1,the intranuclear Foxp3 protein and mRNA expressions of splenic Tregs in mice were markedly down-regulated in 24 h to 72 h(P

Objective To observe temporal expression of matrix metalloproteinases(MMP-13) and tissue inhibitor of metalloproteina-ses-1 (TIMP-1) in lung of newborn rats with hyperoxia induced chronic lung disease (CLD),and to explore the relationship of CLD with MMPs.Methods The neonatal rats within 24 hours after birth were randomly divided into hyperoxia-exposed group(n=40) and control group(n=40).On postnatal 1,3,7,14 and 21 days,lung tissue of rats in 2 groups were collected.Lung histological changes were evaluated by hematoxylin and eosin and Masson stain;Collagen Ⅰ was detected by enzyme linked immunoadsorbent assay;MMP-13 and TIMP-1 were identifide by immunohistochemistry.Results Exposured to hyperoxia enviroment for 21 days,the number of alveolar decreased,terminal air space enlarged,inter-alveolar septa thickened,and deposition of interstitial collagen fibers.On 14 and 21 days,collagen Ⅰ in the lung of hyperoxia-exposed group increased significantly compared with that of control group(P0.05),obviously decreased on 21 day(P

AIM: To survey the pathogenesis, clinical characteristics and surgical treatment for the correction of A-pattern exotropia.METHODS: The case history and surgical effect of 62 A-pattern exotropia in 1825 strabismus were studied. The superior oblique muscles weakening or the horizontal muscles procedures were performed in all patients according to the dysfunction of the superior oblique muscles, horizontal deviation, with or without binocular vision. Follow-up after surgery was 6-18months with mean duration of 9 months.RESULTS: A-pattern exotropia accounts for 3.4% of all strabismus. A-pattern exotropia accounts for 21.2% of A-V syndrome. After surgery, 52 cases got alignment, 56 cases gained A-sign correction and 54 cases recovered binocular vision.CONCLUSION: A-pattern exotropia is a common type in strabismus, its etiology is mainly oblique muscle dysfunction.Different surgical procedures should be selected according to overaction degrees of oblique muscle and binocular vision function, which can acquire the satisfied results.