Imaging study on the homing of regulatory T cells in humanized heterologous liver tissue transplan-tation mouse model / 中华核医学与分子影像杂志

Objective To explore the homing of DiR labeled regulatory T cells ( Tregs) in human-ized heterologous liver tissue transplantation mouse model. Methods The fluorescence intensities of Tregs labeled with different concentrations of DiR dye and different incubation times were measured, and the cell viability was measured by 3-( 4, 5-dimethylthiazol-2-yl )-5-( 3-carboxymethoxyphenyl )-2-( 4-sulfophenyl )-2H-tetrazolium ( MTS) assay to determine the optimal incubation time and dye concentration. The effect of DiR dye on the function and phenotype of Tregs was verified by flow cytometry. The xenogeneic liver tissue transplantation mouse model was constructed and the immune system was reconstituted. Small animal fluores-cence imaging was performed at different time points after infusion of Tregs. Immunohistochemistry analysis was used to analyze immune reconstitution and lymphocyte distribution in vivo. One-way analysis of variance and Dunnett-t test were used to analyze the data. Results With the increase of DiR concentration and incu-bation time, the fluorescence intensity of Tregs increased and gradually weakened after reaching the peak at 3 d. The cell viability of the 5. 00 μg/ml and 20. 00 μg/ml groups was significantly lower than that of the control group (culture medium) at various time points (F=120.142-182.025, t=9.969-19.329, all P<0. 05) . After incubation for 30 min and 60 min, the activity of Tregs was also significantly lower than that of the control group (F=21.826-301.968, t=6.897-40.016, all P<0.05). Tregs were finally co-incubated with DiR dye at a concentration of 2.50 μg/ml for 5 min, which was used further in vivo experiments. The flow cytometry showed that DiR dye did not affect the phenotype or the function of Tregs. The small animal fluorescence imaging showed that Tregs could locate in the graft area of mouse model. Immunohistochemical analysis showed that Tregs could improve lymphocyte infiltration induced by immune reconstitution. Conclu-sion After labeling Tregs with DiR dye, the distribution of Tregs can be directly observed by fluorescence imaging, which is a promising imaging method for Tregs tracer.

Changes of serum Hcy,M-CSF,UA,platelet related indexes and trace element levels in CHD patients with angina pectoris / 心血管康复医学杂志

Objective:To observe the change of serum homocysteine (Hcy),macrophage colony stimulating factor (M-CSF),uric acid (UA),platelet related indexes and trace element levels in coronary heart disease (CHD)pa-tients with angina pectoris.Methods:A total of 58 CHD patients with angina pectoris treated in our hospital from Oct 2010 to Oct 2013 were selected as angina pectoris group,another 58 healthy subjects in same period were en-rolled as healthy control group.Levels of Hcy,M-CSF,UA,platelet related indexes [platelet distribution width (PDW),CD61,CD63]and trace elements were compared between angina pectoris group and healthy control group.Results:Compared with healthy control group,there were significant rise in serum levels of platelet related indexes,Hcy,M-CSF,UA,Zn and Fe,and significant reduction in serum Mn level in angina pectoris group,P <0.05 all;compared with patients with stable angina pectoris,there were significant rise in serum levels of PDW [(13.37±1.61)vs.(17.86±1.99)],CD61 [(45.26±5.11)vs.(58.40±7.20)],CD63 [(14.05±1.96)vs. (18.78±2.95)],Hcy [(13.47±1.52)mmol/L vs.(21.96±2.23)mmol/L],M-CSF [(289.73±31.68)pg/ml vs. (401.56±48.96)pg/ml],UA [(287.95±36.49)μmol/L vs.(373.45±45.26)μmol/L],Zn [(15.76± 1.84)μmol/L vs.(19.55±1.98)μmol/L]and Fe [(12.68±1.50)mmol/L vs.(15.71±1.83)mmol/L],and significant reduction in serum Mn level [(2.21±0.34)μmol/L vs.(1.60±0.21)μmol/L]in patients with unstable angina pec-toris (P <0.05 all).Along with number of coronary diseased vessels increased,serum levels of above indexes signifi-cantly rose except serum Mn level,P <0.05 all.Conclusion:The serum Hcy,M-CSF,UA,platelet related indexes and most trace elements levels significantly rise in CHD patients with angina pectoris,and the abnormal degree is greatly influenced by the angina pectoris type and number of coronary diseased vessels.

Prokaryotic expression and characterization of truncated mutant capsid protein of genotype IV hepatitis E virus / 生物工程学报

A truncated mutant of the Open Reading Frame 2 (ORF2, aa384-606) was amplified from cDNA of genotype IV hepatitis E virus (HEV) by polymerase chain reaction (PCR), subcloned to expression plasmid pTO-T7, and expressed in Escherichia coli. SDS-PAGE and Western blotting were used to detect and identify the recombinant protein, namely rP24. After washing of inclusion bodies, dissolving in denaturing agents, refoldeding by dialysis, ion exchange chromatography and gel chromatography, dynamic light scatter was used to study the hydrated radius of rP24. Western blotting was applied to detect the immunoreactivity of rP24, and mouse immunity test and indirect enzyme linked immunosorbent assay (ELISA) were applied to evaluate the immunogenicity and the detection rate of HEV positive and negative serum. SDS-PAGE and Western blotting show that rP24 was highly expressed in the form of inclusion bodies after induction, and had strong immunoreactivity to monoclonal antibody (McAb) 15B2. After a multi-step purification of rP24, Western blotting indicated that the purified rP24 also had strong immunoreactivity to neutralizing McAb 8C11 and HEV positive serum, suggesting that rP24 simulated the nature structure of HEV capsid protein. Dynamic light scatter demonstrated that the average hydration radius of purified rP24 was 7.48 nm. The mouse immunity test showed that the purified rP24 also had good immunogenicity, and the period of serum antibodies converted from negative to positive was very short, but the antibodies maintained more than 20 weeks. Indirect ELISA tests showed that the detection rate of was the same as anti-HEV-IgG diagnostic kit (Wan Tai corporation). Taken together, the rP24 simulated the neutralizing epitopes of natural HEV, and had strong immunoreactivity and immunogenicity. It provided a basis for the further investigation of the difference of infection mechanism between genotype I and genotype IV HEV.

Effect of San-huang-sheng-fu oil on wounds of full-thickness scald in rabbits / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the effect of San-huang-sheng-fu oil on wounds of full-thickness scald in rabbits.</p><p><b>METHODS</b>Full-thickness scald wounds with area of 6 cm(2) were reproduced on both sides of the back in 9 experimental rabbits by water vapor. These rabbits were divided into sesame oil (S1), San-huang-sheng-fu oil (S2), and mupirocin ointment (M) groups according to the random number table, with 3 rabbits (6 wounds) in each group. Two wounds of each rabbit in the three groups were respectively treated with sesame oil, San-huang-sheng-fu oil, and mupirocin ointment, in a dose of 0.15 mL/cm(2), 2-3 times per day. The general condition of wounds was observed on post scald day (PSD) 1, 11, 22, and 45. The wound healing time was recorded. The wound healing rate was calculated on PSD 5, 11, 15, and 22. All the rabbits were sacrificed on PSD 45, and wound tissues were subjected to histomorphological study with HE staining. The protein expressions of transforming growth factor β1 (TGF-β1), basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) were observed with immunofluorescence staining for the other part of wound tissues. Data were processed with one-way analysis of variance or LSD-t test.</p><p><b>RESULTS</b>(1) The wound healing quality of rabbits in S2 group was better than that in the other two groups. (2) The wound healing time of rabbits in S2 group [(11.2 ± 2.3) d] was significantly shorter than that in S1 group [(21.2 ± 3.1) d, t = 2.591, P < 0.05]. (3) The wound healing rate of rabbit in each group was increased gradually on PSD 5-22. The wound healing rates of rabbits in S2 group on PSD 5-22 were significantly higher than those in S1 group (with t values from 3.920 to 8.605, P values all below 0.05). (4) Histomorphological observation showed that the structure of wound tissues in S2 group was in much better integrity than that in the other two groups, including regenerated hair follicles in the corium layer and regularly arranged collagen fibers. The protein expressions of TGF-β1, bFGF, and VEGF in S2 group were all higher than those in the other two groups.</p><p><b>CONCLUSIONS</b>San-huang-sheng-fu oil can up-regulate the protein expressions of TGF-β1, bFGF, and VEGF, induce vascular regeneration, promote wound healing, and shorten wound healing time.</p>

Comparison of allelic frequencies of 15 short tandem repeat loci between chronic myeloid leukemia patients and non-related healthy individuals / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To compare allelic frequencies of 15 short tandem repeat (STR) loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818 and FGA) between chronic myeloid leukemia (CML) patients and non-related healthy individuals from Changzhou region in order to predict genes related with the CML.</p><p><b>METHODS</b>Blood samples were collected from 745 healthy subjects and 132 CML patients with complete remission. Genotypes were determined with gene scan technology and multiplex PCR with fluorescence-labeled primers. Allelic polymorphisms of 15 STR loci were compared between the two groups. Potential genes related with CML were predicted with statistical analysis of differences in allelic frequencies.</p><p><b>RESULTS</b>Allelic frequencies of 3 loci, including CSF1PO, vWA and TPOX, showed a significant difference (P<0.05) between the two groups.</p><p><b>CONCLUSION</b>CSF1PO, vWA and TPOX loci may be related with CML, albeit that the exact biologic mechanisms is unclear.</p>

The observation of tear ferning in conjunctivochalasis / 眼科

Objective To evaluate tear ferning changes of conjunctivochalasis.Design Prospective case study series.Partici- pants 30 patients(60 eyes)of conjunctivochalasis and normal subjects were selected.Methods The subjects were observed with gen- eral ophthalmic examination and tear fern test(TFT).Tear ferning was classified into 4 types.TypeⅠand TypeⅡare normal.TypeⅢand TypeⅣare abnormal.Main Outcome Measures The type of tear feming.Results TFT showed that tear ferning was de- creased in conjunctivochalasis group(TypeⅢand TypeⅣoccupied 61.7%).The difference between conjunctivoehalasis and normal control group was significant(P