BACKGROUND: The glycoprotein which ends of a-galactosyi residues (a-Gal) is the major heterogenic antigen for hyperacute rejection. OBJECTIVE: To observe the distribution characteristic of the small tissue engineered vessel scaffold, and a-Gal in endothelial cells and smooth muscle cells of Wistar rats and Japanese white rabbits, in addition, to discuss the feasibility of applying acellular tissue vessel scaffold to heterogeneous blood vessel transplantation. DESIGN, TIME AND SETTING: The contrast observation was conducted at the Department of Human Anatomy of China Medical University between March 2003 and December 2004. MATERIALS: Totally 30 caudal arteries of Wistar rats were collected. Fifteen of which were prepared for small vessel scaffold (small vessel scaffold group), the remained 15 served as caudal artery group. Additionally, 15 central arteries were obtained from each ear of Japanese white rabbits (central artery group). METHODS: Totally 16 mg/L Bandeiraea Simplicifolia I Isolectin B4 (BSI-B4) was added for DAB staining with Affinity histochemistry method. Then MetaMorth/C5050/BX41 microscopic image analysis system was used to detect the positive reaction product of a-Gal. MAIN OUTCOME MEASURES: Color changes of vascular wall were observed under light microscope; the optical density of the positive reaction product of a-Gal was measured. RESULTS: The expression of a-Gal was mainly located in the cell membrane, as well as cell nucleus of endothelial cells in the central artery group. The expression of a-Gal of endothelial cell was strong positive in the caudal artery group, which was weak or negative expressed in the small vessel scaffold group. The optical density of a-Gal expression was lowest in the tunica intima of small vessel scaffold group, which was less in the central artery group than the caudal artery group (P < 0.001). The a-Gal expression in the tunica media of small vessel scaffold group was less than the central artery and the caudal artery groups (P < 0.001). CONCLUSION: In the caudal artery of Wistar rat, the expression of a-Gal is higher than that in the central artery of Japanese white rabbit. Therefore, the heterogeneous tissue engineered vessel material from acellular caudal artery of Wistar rats can be used in blood vessel transplantation.

Objective To investigate the changes of follicular regulatory T cells ( Tfr cells) and follicular T helper cells ( Tfh cells) in peripheral blood of children with myasthenia gravis ( MG) . Methods We recruited 28 MG patients and 20 healthy subjects in this study. The percentages of Tfh and Tfr cells in peripheral blood samples were measured by flow cytometry. Real-time PCR was performed to detect the ex-pression of transcription factors and regulatory factors of Bcl-6, c-MAF, Blimp-1 and PD-1 at mRNA level. ELISA was used to detect the levels of IL-2, IL-6, IL-10 and IL-21 in plasma samples and the titers of Ach-Rab and PsMab. Results Compared with the healthy subjects, the MG patients showed higher percentages of Tfh cells and lower percentages of Tfr cells before receiving treatment. The expression of Bcl-6 and c-MAF on CD4+T lymphocytes cells at transcriptional level were significantly enhanced, while the expression of Blimp-1 on CD4+T cells and the expression of PD-1 on Treg cells at transcriptional level were inhibited in the MG patients in comparison with those in healthy subjects. Moreover, decreased levels of IL-2 and increased levels of IL-21 were found in plasma samples collected from the MG patients. Conclusion The decreased percentages of Tfr cells and increased percentages of Tfh cells in patients with MG resulted in abnormal ratios of Tfr/Tfh cells, which might be involved in the immunological pathogenesis of MG. Several changes in the patients with MG might be responsible for the imbalanced ratio of Tfr/Tfh cells, which included changes of IL-2 and IL-21 in microenvironment, enhanced expression of Bcl-6 and c-MAF at mRNA level and inhibited expression of Blimp-1 at mRNA level on CD4+T cells as well as over-expression of PD-1 at mRNA level on Treg cells.

Objective To investigate the influence of IVIG on immunologic function and cytokines levels in children with EV71 infection associated high-risk pulmonary hemorrhage. Methods According to the inclu-sion criteria , 64 children were enrolled and randomly divided into two groups: 39 cases in the IVIG treatment group and 25 cases in the general treatment group. The alternations of blood and immune cytokine markers before and after treatment were detected in the patients. Results (1) Before treatment, the peripheral blood T cells, TH and B cells in the IVIG group were higher than those in the general group , but the peripheral blood IgA was lower than that in the general group(P < 0.05); (2) In the IVIG group, the NK cells and IgG increased and T cells, TH cell and B cells significantly decreased after receiving IVIG therapy (P < 0.05); (3)No significant difference was observed in the levels of IL-6 and IL-10 in the two treatment groups before treatment. However , after IVIG treatment, IL-6 and IFN-γ levels reduced, IL-10 level increased (P < 0.05). After the general treat-ment, no significant difference was found in levels of IL-6,IL-10 and IFN-γ in peripheral blood (P > 0.05). Conclusion Disorders of cellular immunity and humoral immunity appeared in children with EV71 infection-re-lated high-risk pulmonary hemorrhage. It has clinical value to use IVIG timely to regulate the immune disorder.

Objective To explore the clinicopathological features and prognostic significance of human epidermal growth factor receptor 2 (HER2) in gastric carcinoma. Methods Pathological data of 127 patients with gastric carcinoma were retrospectively analyzed. HER2 expressions of all patients were detected by immunohistochemistry (IHC). 119 (93.7 %) patients were undergone R0 dissection and 123 (96.9 %) cases received D2 lymph nodes dissection. 51 (40.2 %) patients received adjuvant chemotherapy. The proportional differences of clinicopathological features for patients between HER2-positive and HER2-negative were compared, including the patients' survival. Results HER2 overexpression rate was 8.7 % (11/127) in gastric carcinoma patients. For the patients with HER2-positive and HER-negative, the lymph node metastasis rates were 100.0 % (11/11) and 81.9 % (95/116), respectively (P= 0.041). The 3-year overall survival (OS) rates for HER2-positive and HER2- negative patients with gastric carcinoma were 32.7 % and 42.9 % (P=0.413), and the 5-year progression free survival (PFS) rates were 27.3 % and 42.2 % (P = 0.354), respectively. Among patients with HER2-negative, 3-year OS rate for patients with surgery plus adjuvant chemotherapy was 55.3%, compared with 35.4%for patients with surgery alone (P=0.015), and the 3-year PFS rates were 53.3 % and 35.3 % (P= 0.038), respectively. Among patients with HER2-positive patients, 3-year OS rate for patients with surgery plus adjuvant chemotherapy was 0, compared with 75.0%for patients with surgery alone (P=0.002), and the 3-year PFS rates were 0 and 60.0% (P=0.004). Conclusions HER2 is expressed in gastric carcinoma tissue, related to lymph node metastasis. HER2 status are not correlated with the prognosis for gastric carcinoma patients, however, it is likely to be a predictive marker for adjuvant treatment after surgery for patients with gastric carcinoma.

Objective To observe the clinical effects of L-carnitine in myocardial enzyme abnormality caused by enterovirus in children with hand foot and mouth disease (HFMD). Methods 660 HFMD children patients with myocardial enzyme abnormality from May 2013 to June 2016 were enrolled and randomly divided into three groups. Group A(n=220)was treated with 1′6-FDP,group B with L-carnitine and group C with L-carnitine combined with 1′6-FDP. All groups were given routine anti-virus and symptomatic treatment. The clinical efficacy was compared across the three groups in terms of myocardial enzyme spectrum,heart rate,ECG,severe conver-sion rate before and after treatment. Results (1)Before treatment,there was no significant difference between the three groups in gender,age,course of disease,heart rate,myocardial enzyme spectrum and other indicators. (2)After treatment,the cure rate of HFMD in group B and group C were significantly higher than that in group A (both P<0.05);the rate of severe cases and the ECG normal rate in group B and group C were significantly lower than those in group A(both P<0.05),the time for heart rate resuming to normal in group B and in group C was all significantly shorter than that in group A(both P<0.05). There were no significant differences between group B and group C in clinical cure rate,severe conversion rate,recovery rate of ECG and heart rate recovery(all P>0.05).(3)In comparison with group A,after treatment,the levels of myocardial enzyme in group A and group B were decreased significantly(P < 0.05)and the recovery rates of myocardial enzyme in group A and the group B were significantly higher (P < 0.05),but no significant difference were observed between group A and group B (P>0.05). Conclusions For HFMD children with myocardial enzyme abnormality caused by enterovirus ,L-car-nitine together with myocardial nutritional therapy can significantly improve the myocardial enzyme indexes and electrocardiogram abnormality. It reduces the rate of severe cases and improve the prognosis.

Objective: To observe the neuro-protective effect of Levocarnitine on severe hand, foot and mouth disease (HFMD) after enterovirus 71(EV71) infection, to preliminarily explore the possible mechanism preliminarily. Methods: One hundred and thirty-two children with EV71 infection and HFMD combined with serum S100 protein and neuronspecific enolase (NSE) abnormalities who were admitted to Children′s Hospital Affiliated to Zhengzhou University from March 2015 to July 2016 were enrolled in the study.They were divided into the routine group and the Levocarnitine group by the random number grouping method.The routine group (66 cases, including 32 males and 34 females, median age of 2 years and 3 months) was given symptomatic treatment such as antiviral therapy while the Levo-carnitine group (66 cases, including 36 males and 30 females, median age of 2 years and 5 months) was treated with Levocarnitine for neuroprotection on the basis of routine group.Forty healthy children (23 males and 17 females, median age of 2 years and 6 months) who were examined at the Children′s Hospital Affiliated to Zhengzhou University during the same period were selected as the healthy control group.The levels of S100, NSE, soluble apoptosis-related factors (sFas), soluble apoptosis-related factor ligands (sFasL), malondialdehyde (MDA), superoxide dismutase (SOD) in serum were compared between the healthy control group and children with HFMD.The levels of above-mentioned indexes in cerebrospinal fluid and serum, efficacy-related indicators such as duration of fever, white blood cell count on the 3^rd day of treatment, time to remission of nervous system symptoms, time of disease progression and critical conversion rate were compared between 2 groups of children with HFMD.The correlation between sFas, sFasL, MDA, SOD and S100, NSE was performed Results: (1) The levels of S100 [(0.38±0.16) μg/L vs. (0.06±0.23) μg/L], NSE [(43.70±8.80) μg/L vs. 10.10±3.60) μg/L], sFas [(6.61±1.86) μg/L vs. (3.88±1.22) μg/L], sFasL[(101.40±20.7) μg/L vs. (54.4±13.3) μg/L] and MDA[(11.98±2.54) nmol/L vs. (4.08±1.45) nmol/L] in serum of HFMD group were significantly higher than those of the healthy control group (t=-12.245, -22.895, -8.273, -12.803, -17.960, all P<0.05), while the SOD level [(57.10±10.40) kU/L vs. (70.3±14.4) kU/L] was significantly lower (t=5.457, P<0.05). (2) With the extension of treatment time for HFMD children in the two groups, S100 and NSE in cerebrospinal fluid, S100, NSE, sFas, sFasL and MDA in serum decreased, while SOD level increased.On the 3^rd and 7^th day after treatment, S100 (t₃=3.491, t₇=14.434), NSE (t₃=2.920, t₇=23.490) in cerebrospinal fluid, S100 (t₃=5.277, t₇=3.614), NSE (t₃=4.652, t₇=10.525), sFas (t₃=6.399, t₇=7.514), sFasL (t₃=11.155, t₇=8.804) and MDA (t₃=6.348, t₇=7.499) in serum of Levocarnitine group were significantly lower than those of routine group (all P<0.05), while SOD (t₃=3.162, t₇=-3.529) was significantly higher than that of routine group (P<0.05). (3) The relief time of neurological symptom in levocarnitine group was significantly shorter than that in the routine group [(1.23±0.65) d vs. (1.84±0.47) d], and WBC on the 3^rd day after treatment [(9.14±2.93)×10⁹/L vs. (7.12±2.58)×10⁹/L] and the progression time of the disease [(29.74±7.85) h vs. (17.36±8.73) h] were significantly better than the those in the routine group (t=-6.178, 4.204, 8.567, all P<0.05). The critical conversion rates of Levocarnitine group and the routine group were 7.58% and 18.18%, respectively, and the difference in critical conversion rate was not statistically significant (χ²=2.316, P>0.05). (4)There was a positive correlation between S100 and sFas, sFasL, MDA in children with HFMD (r=0.373, 0.735, 0.334, P<0.05). NSE was positively correlated with sFas and sFasL (r=0.479, 0.601, all P<0.05), while SOD and S100 were negatively correlated with NSE (r=-0.425, -0.460, all P<0.05). Conclusions Levocarnitine has good curative effect on severe HFMD in children infected by enterovirus EV71, which can effectively protect the cranial nerves.The mechanism may be related to scavenging oxygen free radicals and blocking nerve cell apoptosis.

Objective:To explore the clinical significance of serum Chromogranin A (CGA) level in predicting the prognosis of children with severe hand, foot, and mouth disease (HFMD) and complicating neurogenic pulmonary edema (NPE).Methods:A total of 162 patients with HFMD admitted in our hospital from January 2017 to December 2019 were enrolled in the study; and 40 age-matched healthy children were selected as controls. According to the disease severity and complication the patients were divided into three groups: mild group ( n=88), severe without NPE group ( n=46) and severe with NPE group ( n=28). In 72 severe HFMD patients 16 cases died (fatal group) and 56 cases survived (survival group) within 28 days of hospitalization. The serum CGA, LAC, GLU, WBC, PCT, IL-6, cTnT were measured in all subjects. SPSS 23.0 software was used for data analysis, and the receiver operating characteristic (ROC) curve was used to evaluate the various indicators for predicting the prognosis of severe HFMD combined with NPE. Results:The serum CGA, GLU, LAC, IL-6 and cTnT levels in severe HFMD group with NPE significantly higher than those in the other three group ( H=61.554, 79.031, 86.994, 36.477, 75.021, all P<0.05 ). The serum CGA, LAC, GLU and IL-6 levels in the fatal group were significantly higher than those in survival group ( Z=-6.094, -4.621, -4.283, -5.504, all P<0.05). There was no significant difference in the levels of WBC, PCT and cTnT between the survival group and the fatal group ( P>0.05). The area under the receiver operating curve (AUC) of serum CGA was 0.890 (95% CI: 0.833-0.947) for predicting the prognosis of patients and the best cut-off value was 120.59 μg/L. Conclusion:The detection of serum CGA levels may be beneficial for the early diagnosis of severe HFMD with NPE, and can be used as one of the predictors of death from severe HFMD.

Objective To explore the protective effect of magnesium sulfate on the nerve injury in severe hand, foot and mouth disease ( HFMD) caused by enterovirus A71 ( EV-A71) and to investigate its clinical and prognostic effects.Methods A total of 240 cases of severe HFMD with EV-A71 infection and nerve injury were enrolled.According to the random number table method, the patients were randomly divided into conventional treatment group (control group) and magnesium sulfate treatment group ( treatment group), with 120 cases in each group.The control group was given the routine treatment, and the treatment group was given the magnesium sulfate adjuvant treatment on the basis of routine treatment.The neurological symptoms and signs, clinical efficacy and prognosis were observed before and after treatment in the two groups.The blood and cerebrospinal fluid neuron-specific enolase ( NSE), S100-βprotein and neuropeptide Y ( NPY) were analyzed before and after treatment.The amplitude integrated electroencephalogram (aEEG) was used to monitor the abnormal recovery of EEG.The t-test was applied to analyze quantitative data, and the chi-square test was used for qualitative data comparison.Results Among children with severe HFMD, there were 83 cured cases, 29 improved cases and 8 ineffective cases in control group, with the total effective rate of 93.3%; while in the treatment group, 101 cases were cured, 18 cases were improved and 1 case was ineffective, the total effective rate was 99.2%.The therapeutic effects (Z＝2.918, P＝0.004) and the total effective rate ( χ2 ＝4.156, P＝0.041) were statistically significantly different between the two groups.Three days after treatment, the average levels of serum NSE, S100-βprotein and NPY in magnesium sulfate treatment group were significantly lower than those in control group (t＝-7.239,-10.020 and -11.053, respectively, all P＜0.01).Five days after treatment, the average levels of cerebrospinal fluid NSE, S100-β protein and NPY in magnesium sulfate treatment group were significantly lower than those in control group ( t＝-6.546,-13.308 and -10.258, respectively , all P＜0.01).After treatment, the neurological function score in treatment group was significantly lower than that in control group and that before treatment , and the differences were statistically significant ( t ＝-9.473 and 12.162, respectively, both P ＜0.01 ).The recovery time of the main symptoms and signs in treatment group was ( 2.33 ±0.76 ) d, which was significantly shorter than that of control group ([3.21 ±0.82] d), the difference was statistically significant (t＝-12.52, P＜0.05).The average length of hospital stay in treatment group was (5.79 ±1.42) d, which was shorter than that in control group ([ 6.71 ±1.46 ] d ), and the difference was statistically significant ( t＝-4.932, P＜0.05).Of the 240 children with severe HFMD, 194 (80.8%) patients had abnormal aEEG.Before treatment, the aEEG abnormal rates in control group and the magnesium sulfate treatment group were 79.2%(95 cases) and 82.5%(99 cases), respectively, there was no significant difference ( χ2 ＝0.430, P＞0.05); while after treatment for 3 days, 76 cases in treatment group returned to normal, and the recovery rate of aEEG was 76.8%, which was higher than that in control group (52.6%). The difference was statistically significant ( χ2 ＝12.406, P ＜0.05 ).Conclusions Magnesium sulfate adjuvant therapy can reduce the abnormal levels of NSE, S100-βand NPY in blood and cerebrospinal fluid, relieve clinical symptoms, shorten the course of disease and average length of hospital stay, improve the neurological function score, and promote the recovery of abnormal aEEG.Thus, it has neuroprotective effect on severe HFMD with nervous system lesion.

Objective To investigate the significance of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in serum and cerebrospinal fluid for evaluation of severe hand ,foot ,and mouth disease (HFMD) complicated with neurogenic pulmonary edema (NPE).Methods A total of 140 patients diagnosed with HFMD in Henan Children′s Hospital were enrolled and divided into three groups including mild group ,severe HFMD group without NPE ,severe HFMD group with NPE .These severe HFMD patients were also divided into survival group and death group according to the 28-day prognosis .Meanwhile ,50 age-matched healthy children were selected as controls .Serum MMP-9 and TIMP-1 levels were measured in all enrolled children .At the same time ,MMP-9 ,TIMP-1 and ratio of MMP-9／TIMP-1 in cerebrospinal fluid were measured in the severe HFMD group with and without NPE .Quantitative data were compared using one-way analysis of variance , and means comparisons between samples were conducted using LSD-t test .Results Among 140 children with HFMD ,66 were in mild group ,42 in severe HFMD without NPE group ,and 32 in severe HFMD with NPE group .And 50 healthy children were in control group .After 28 days ,14 cases died in severe HFMD groups .MMP-9 , TIMP-1 and MMP-9／TIMP-1 in serum of severe HFMD group with NPE increased significantly greater than those in the other three groups (F＝269 .356 ,121 .301 and 101 .502 ,respectively ,all P ＜0 .05). MMP-9 ,TIMP-1 and MMP-9／TIMP-1 in cerebrospinal fluid of severe HFMD group with NPE were (57 .24 ± 8 .92) μg／L ,(35 .26 ± 8 .14) μg／L and (1 .66 ± 0 .23) μg／L ,respectively ,while those in cerebrospinal fluid of severe HFMD group without NPE were (30 .57 ± 3 .89) μg／L ,(26 .25 ± 0 .32) μg／L and (1 .17 ± 0 .61) μg／L ,respectively .The differences between the two groups were all statistically significant (t＝62 .485 ,37 .680 and 169 .387 ,respectively ,all P＜0 .01).MMP-9 ,TIMP-1 and MMP-9／TIMP-1 in serum and cerebrospinal fluid of death group increased significantly greater than those in survival group ,the difference were statistically significant (all P＜0 .01).The maximum area under curve (AUC) was reached when the MMP9／TIMP-1 ratio in cerebrospinal fluid was 0 .890 (95% CI :0 .801 －0 .978).Conclusions MMP-9 and TIMP-1 may be involved in the pathogenesis of HFMD complicated with NPE .The detection of MMP-9 and TIMP-1 levels may be beneficial for the early diagnosis of severe HFMD with NPE .The imbalance of MMP-9／TIMP-1 ratio can be used as one of the predictors of severe HFMD combined with NPE.

Objective: To investigate the value of abnormal expression of HLA-DR on peripheral blood monocytes in evaluating the immune function status, clinical prognosis and severity of patients with hand, foot and mouth disease (HFMD). Methods: From June 2017 to October 2018, 100 cases of mild HFMD, 80 cases of severe HFMD, 32 cases of critical HFMD and 40 healthy children (control group) were recruited in this study. The patients were divided into two groups, lower DR group (DR-L, HLA-DR expression<30%) and normal DR group (DR-N, HLA-DR expression>30%) according to the HLA-DR expression on monocytes. Flow cytometry was used to detect the CD14⁺ monocytes expressing HLA-DR and the absolute count of lymphocyte subsets. Immunoturbidimetry was used to detect the levels of IgG, IgM and IgA in plasma samples. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the levels of IFN-γ and IL-10 in plasma samples. Pediatric critical illness score (PCIS) and the pediatric risk of mortality Ⅲ (PRISM Ⅲ) were used to estimate the severity of HFMD. Results: ① There were significant differences in HLA-DR expression on monocytes among children with mild, severe and critical HFMD (F=47.102, P<0.05). Patients with critical HFMD had the lowest HLA-DR expression (P<0.05). ② The numbers of CD14⁺ monocytes, CD3⁺ T cells, CD4⁺ T cells, CD8⁺ T cells, B cells and NK cells in peripheral blood of the DR-L group were significantly lower than those of the DR-N group and the normal group, especially in patients with severe or critical HFMD (P<0.05). ③ There was no significant difference in the level of IgG, IgA or IgM among the DR-L, DR-N and control groups (P>0.05). ④ Compared with the DR-N group, the DR-L group showed decreased IFN-γ level and increased IL-10 level in plasma (P<0.05). The ratio of IFN-γ/IL-10 of the DR-L group was lower than that of the DR-N group and control group (P<0.05). HLA-DR expression was negatively correlated with the concentration of IL-10 in plasma (r=-0.704, P<0.05), and positively correlated with the IFN-γ/IL-10 ratio (r=0.773, P<0.05). ⑤ Compared with the DR-N group, the DR-L group showed lower PCIS and higher PRISM Ⅲ. HLA-DR expression was positively correlated with PCIS (r=0.715, P=0.00) and negatively correlated with PRISM Ⅲ (r=-0.610, P=0.00). ⑥ The incidence of pulmonary edema, pulmonary hemorrhage and cardiopulmonary failure and the mortality of HFMD patients in the DR-L group were significantly higher than those in the DR-N group (P<0.05). Conclusions Patients with severe or critical HFMD had cellular immune dysfunction and abnormal HLA-DR expression on CD14⁺ monocytes. Assessing the expression of HLA-DR on monocytes could be used to evaluate the cellular immunity of patients with severe or critical HFMD. Lower expression of HLA-DR on CD14⁺ monocytes might be associated with severe HFMD and poor prognosis.