Previous work showed that there was high ratio of Mycoplasma hyorhinis infection in human gastric carcinoma. P37 protein is a membrane lipoprotein of Mycoplasma hyorhinis. It was reported that P37 inhibited cell adhesion and induced tumor invasiveness. To investigate the function of P37 in cancer development, the p37 gene was subcloned into shuttle vector of pAdTrack-CMV. Linearized pAdTrack-CMV-p37 was co-transformed into BJ5183 cells with adenoviral genomic plasmid pAdEasy-1. The identified recombinant DNA was transfected into 293 cells to package adenovirus. From the supernatant and cell lysis, the presence of recombinant adenovirus P37 was proved by PCR. And then BICR cells with this recombinant adenovirus of P37 were successfully infected. RT-PCR and Western blot demonstrated the transcription and expression of P37 in infected BICR cells. And the soluble P37 protein can be secreted into the culture supernatant of infected BICR cells. In the migration assay in vitro, the number of migration BICR cells infected with Ad-p37 was 52.03% more than that of control. The construction of recombinant adenovirus provided a good tool for studying P37 function and its molecular mechanism, which will be further used for in vivo migration and invasion experiments.

Objective:To research the number and function of monocyte-macrophages in patients with chronic active hepatitis B. Methods:The 51 chronic viral hepatitis B( CHB) patients were selected randomly,which consisted of 20 cases of mild-moderate,31 cases of severe group and 13 cases of healthy controls. PBMCs were separated by percoll. Monocytes were tagged by CD14,the molecules CD80,CD86,HLA-DR and CD163 were detected by flow cytometry which expressed on the surface of PBMCs. Serum cytokine were detected for IL-10, IL-12 and IL-23 by ELISA. The distribution of CD68 was detected in the liver by immunohistochemical staining. Results:The expressions of CD80 for all chronic hepatitis B patients were lower than the controls respectively,no matter mild-moderate or even severe group. Similarly,the HBV patients expressed lower level of CD86 in the peripheral blood mononuclear cells when compared with the control group. Furthermore, there was statistically difference between the levels of CD86 in severe group compared with control group (P<0. 01). As the expression of CD80 and CD86,the levels of HLA-DR in the patents had also declined when compared with controls. While the HLA-DR levels in both the mild-moderate HBV hepatitis groups were statistically significant higher than the severe group (P<0. 01). Different from the above all,the expression of CD163 in all chronic HBV hepatitis was higher than the control group. The CD68 positive cells in chronic HBV patients were observed and infiltrated increasingly in portal area and hepatic lobules (P<0. 05). There were statistically significant differences of IL-10 levels between the mild-moderate group,severe group and the control group,respectively (P<0. 01). Conclusion:Macrophages have participated in the pathological lesions of liver in CHB patients,among peripheral blood mononuclear cells,the phenomena of imbalance between type M1/M2 and polarization to type M2 have been observed,which participated in the development of the chronicity of CHB.

Objective To evaluate the changes of serum tumor markers of carbohydrate antigen 125 (CA125),carbohydrate antigen 199 (CA199),carcinoembryonic antigen (CEA) and squamous cell carcinoma antigen (SCC) during anti-tuberculosis treatment in patients with active pulmonary tuberculosis and tuberculous pleurisy.Methods This research was a prospective study.Sixty-three patients with active pulmonary tuberculosis and 24 patients with tuberculous pleurisy underwent blood samplings before treatment,2 and 6 months after treatment.Centaur XP chemiluminescent immunoassay was used to test serum levels of CA125,CA199,CEA and SCC.Thirty healthy subjects were included as controls.Student t-test was used to compare continuous variables,and chi-square test or Fisher exact test was used to compare categorical variables.Serial changes of serum tumor markers levels pre-and post-treatment were analyzed by repeated measures analysis of variance.Binary Logistic regression model was used for multivariate analysis.Results The mean serum CA125 level of the 63 patients with active pulmonary tuberculosis pre-treatment was (64.4± 30.4) U/mL,which was significantly higher than that of healthy controls ([12.7±5.5] U/mL,t=11.98,P＜0.01).The mean serum CA125 level decreased to (16.9±6.1) U/mL after 6 months standardized treatment,which was significantly lower than that before treatment (t=12.74,P＜0.01).While compared with healthy controls,the serum level of CA125 in patients who had completed the standardized treatment was not significantly different (t =0.94,P =0.348).When compared with the healthy controls,serum CA199,CEA,SCC levels in patients with active pulmonary tuberculosis before and after treatment showed no statistically significant difference (P＞ 0.05).Univariate analysis and multivariate analysis by Logistic regression analysis showed that bilateral pulmonary tuberculosis (x2 =7.746,P=0.006; OR=6.99,95％CI:1.73-28.22) and cavity pulmonary tuberculosis (x2 =6.254,P=0.012; OR=7.64,95％CI:1.64-35.35) were associated with increased serum CA125 level.The mean serum CA125 level of 24 cases of tuberculous pleurisy pre-treatment was (81.2 ± 37.6) U/mL,which was both significantly higher than that of patients with active pulmonary tuberculosis (t=2.153,P=0.034) and that of healthy controls (t=12.05,P＜0.01).Similarly,CA125 levels sharply decreased to (15.5 ± 7.3) U/mL after 6 months standardized treatment,which was not statistically significant compared with the control group (t=0.450,P=0.652).However,CA199,CEA and SCC levels in tuberculous pleurisy pre-and post-treatment were all not statistically different from those of healthy controls (all P＞0.05).Conclusion Serum CA125 can be used as a marker for assessing the disease progression and therapeutic efficacy for patients with active pulmonary tuberculosis and tuberculous pleurisy.

Objective:To study the preparation and quality control of loxoprofen sodium patches and develop an HPLC method for the determination of loxoprofen sodium. Methods:Loxoprofen sodium patches were prepared with CMC-Na and PVP as the adjuvants, and an HPLC method was used to determine the content of loxoprofen sodium. Results:The linear range of loxoprofen sodium was 4-24μg·ml-1(r=0. 999 7), the average recovery was 99. 99%(RSD=0. 99%, n=6). Conclusion:The preparation method is reason-able, simple and stable. The developed HPLC method is specific to determine the content of loxoprofen.

Objective Glucose metabolism trend was dynamicly mornitored following liver transplantation, and its affecting factors were assessed. Methods The glucose metabolism status were assessed at four time points respectively after liver transplants, then they were divided into two groups:normal glucose metabolism (NGM) and abnormal glucose metabolism (AGM). The clinical data were univariate analyzed and multivariate analyzed to screen the risk factors. Results At 1 month, 3 months, 1 year and 3 years post-transplantation, the incidence of AGM were 74.0%, 43.9%, 29.4%, 24.1% respectively Between these two groups, age > 45 y had a significant difference at 1 month, 3 months, 1 year and 3years post-transplantation; the use of tacrolimus had a significant difference at 3 months, 1 year and 3years post-transplantation, but the dose of tacrolimus or tacrolimus blood concentration showed no significant difference; high dose of glucocorticoid had significant difference at 1 month , 3 months post-transplantation; high BMI and acute rejection had significant difference at 1 month post-transplantation. Conclusions There is a high incidence of abnormal glucose metabolism (AGM) in the early stage post-transplantation, and a considerable number of patients' glucose metabolism improved in the later period. Age>45 y and tacrolimus affect glucose metabolism for a longer period post-transplants. High BMI and acute rejection have an impact on glucose metabolism only in the early stage post-transplantation. Large dose of glucocorticoid affect glucose metabolism for at least 3 months post-transplantation , and there is no significant difference after 1 year.

Objective To systematically review the effects of different delivery routes of oxytocin in preventing hemorrhage after cesarean section.Methods Databases including PubMed,Medline,Embase,Cochrane library,Wanfang Medical Database,China National Knowledge Internet (CNKI),VIP Database for Chinese Technical Periodicals were searched by computers and Conference papers were manually searched.The randomized,controlled clinical trials of oxytocin given by intramuscular injection,intravenous injection or intravenous infusion were included in elective cesarean section under spinal anesthesia.The quality of included literatures was evaluated by Cochrane systematic evaluation.The primary outcome measure was intraoperative amount of blood loss or change in Hb before and after operation.The secondary outcome measures were changes in blood pressure and heart rate,electrocardiogram,uterine contraction and other adverse reactions after using oxytocin.Results Seven studies involving 2 325 patients were included in this meta-analysis.Among the 2 325 patients,oxytocin was given through intramuscular injection in 79 cases,by intravenous injection in 1 147 cases and in the way of intravenous infusion in 1 099 cases.Compared with intramuscular injection group or intravenous infusion group,no significant change was found in the amount of blood loss during operation in intravenous injection group (P＞0.05).There was no statistically significant difference in the amount of blood loss during operation between intravenous injection group and intravenous infusion group (P＞0.05).Intravenously infusing oxytocin produced less effect on the mean arterial pressure and heart rate than intravenously injecting oxytocin (P＜0.01).Conclusion Intravenous infusion is a suitable route for oxytocin delivery in cesarean section with spinal anesthesia.

Objective To study the effects of thymus transplantation(TT)combined with CD4--DLI on T cell reconstitution after allogeneic hematopoietic stem cell transplantation(allo-HSCT). Methods BALB/c mice were randomly divided into three groups:hematopoietic stem cell transplantation (HSCT group),hematopoietic stem cell transplantation combined with thymus transplantation(TT group)and hematopoietic stem cell transplanta-tion combined with thymus transplantation plus CD4+ T cell-depleted lymphocyte infusion(CD4--DLI group). On day-1,the mice were treated with the lethal dose of radiotherapy. On day 0,C57BL/6 mice were used as donor for hematopoietic stem cell transplantation. The mice were sacrificed on 5 days,2 weeks,4 weeks and 3 months after transplantation,respectively. The peripheral blood and spleen cells of mice were collected for determinations of T cell surface antigen,T cell receptor,naive T cells and intracellular cytokines. HE staining was used to assess the development of donor thymus. Results TT and CD4--DLI did not impair each other′s effects on T cell reconstitu-tion. TT combined with CD4--DLI increased the number of T cell reconstruction. CD4--DLI promoted the effect of TT on enlargement naive CD4+and CD8+T cell pool. Combination of TT and CD4--DLI enhanced the cytokine pro-duction of T cells. Conclusion TT combined with CD4--DLI had no side effects on TCR repertoire and thymus. Conclusion TT combined with CD4--DLI can enhance the reconstitution of T cell number and function via thymus dependent and thymus independent mechanism.

Objective To investigate the feasibility and safety of surgical device closure of doubly committed sub-arterial ventricular septal defect via left sub-axillary.Methods A total of 45 patients diagnosed as doubly committed sub-arterial ventricular septal defect (dcVSD) with transthoracic echocardiography (TTE) and transesophageal echocardiography(TEE) were enrolled from June 2014 to August 2016 in Henan Children Heart Center,Henan Provincial People's Hospital.There were 39 males and 6 females,with the mean age of (2.2 ±2.1) years old(0.5-8.0 years),the body weight (13.8 ± 7.1) kg(7.0-34.1 kg),the defect size (4.5 ± 1.0) mm (3.0-8.0 mm).After general anesthesia,the patients were in supine and evaluated by TEE which indicated whether they were fit to closure.Then,they were turned to the right lateral position while this technique was determined.A vertical incision of 2-3 cm was made between the third and the fifth intercostal space and invasion in thoracic space via fourth intercostal space.Puncture was done at the anterior surface of right ventricular outlet tract to build a delivery tract.The occluder was released and the VSD was occluded under transesophageal echocardiography guidance.Results Forty-one patients had a successful surgical dcVSD closure with asymmetric occluders sized (6.0 ± 1.5) mm(4-10 mm).Among 4 failure cases,2 cases (4.4％) were switched to open-heart surgical repair,1 case (2.2％) due to device related aortic regurgitation,the rest 1 case (2.2％) experienced a dislocation of occluder into pulmonary artery and was converted to surgical repair after retrieve of occluder.Trivial residual shunt was detected in 2 cases (4.4％) postoperatively,a spontaneous closure was observed by 1 month follow-up and 3 months follow-ups,respectively.All the patients were discharged 5 to 8 days after the operation.With a follow-up of (10.4 ±5.0) months [3-24 months],there were no complications such as pericardial effusion,displacement of device,atrioventricular block or new valvular dysfunction.Conclusions Minimally invasive device closure of doubly committed sub-arterial ventricular septal defect via left sub-axillary is a feasible and safe treatment for closure of dcVSD.This technique has advantages of minor wound,less exudation,covert incision,however,long term follow-up is necessary.

Objective: To investigate the changes and clinical significance of vascular endothelial (VE)-cadherin and procalcitonin (PCT) in serum and cerebrospinal fluid (CSF) of children with viral encephalitis or bacterial meningitis(BM). Methods: A total of 42 cases of children with viral encephalitis(viral encephalitis group), 36 cases of children with BM(BM group), and 20 cases of children with non-nervous system injury(control group) were selected from September 2016 to June 2018 at the Third Hospital of Zhengzhou University.The serum and CSF levels of VE-cadherin and PCT levels of the 3 groups were detected by using enzyme-linked immunosorbent assay. Results: The levels of VE-cadherin in the serum of viral encephalitis group, BM group and control group at the acute phase were (5.60±1.17) mg/L, (7.08±1.01) mg/L and (2.52±0.68) mg/L respectively, and the levels of VE-cadherin in CSF of viral encephalitis group, BM group and control group were (6.00±1.09) mg/L, (6.97±1.11) mg/L and(1.93±0.88) mg/L, respectively.The levels of PCT in the serum of viral encephalitis group, BM group and control group at the acute phase were (0.26±0.11) μg/L, (0.82±0.17) μg/L and (0.27±0.13) μg/L, respectively, and the levels of PCT in the CSF of viral encephalitis group, BM group and control group were (0.25±0.11) μg/L, (0.72±0.14) μg/L, (0.28±0.17) μg/L, respectively.As a result, the levels of VE-cadherin and PCT in the serum and CSF of BM group showed significant increase, compared with viral encephalitis group and control group in the acute phase(F=124.94, 163.21, 151.62, 127.37, all P<0.01). The levels of VE-cadherin in the serum and CSF of viral encephalitis group were also significantly higher than that of control group (all P<0.01), but there was no difference between viral encephalitis group and control group about the levels of PCT in the serum and CSF (all P>0.05). The levels of VE-cadherin in the serum of viral encephalitis group and BM group after treatment were (2.34±0.81) mg/L and (2.67±1.29) mg/L, and were(2.55±0.92) mg/L and(2.39±0.74) mg/L in the CSF.The levels of PCT in the serum of viral encephalitis group and BM group after treatment were (0.25±0.11) μg/L, (0.30±0.17) μg/L, and the levels of PCT in the CSF of viral encephalitis group and BM group were(0.22±0.10) μg/L and (0.27±0.12) μg/L.After effective treatment, the levels of VE-cadherin, PCT in serum and CSF of BM group and viral encephalitis group were almost equal, and the difference was not statistically significant(F=1.83, 0.76, 2.72, 3.89, all P>0.05). In the receiver operating characteristic (ROC) curve, the area under the ROC curve of VE-cadherin in serum and CSF was 0.896 and 0.912, and was 0.670 and 0.668 of PCT respectively. Conclusions VE-cadherin may be involved in the early stage of intracranial infection, and it may be helpful in differentiation of virus or bacterial infection with PCT.VE-cadherin has a good diagnostic value for intracranial infection.

Objective:To explore the effects of apatinib on the proliferation and apoptosis of FLT3-ITD mutant acute myeloid leukemia (AML) cells, and to explore the related mechanisms.Methods:The logarithmic growth phase FLT3-ITD mutant AML cell lines MV4-11 and MOLM-13 were treated with different concentration of apatinib for 48 hours. The cell proliferation was detected by CCK-8 method. Flow cytometry was performed to examine the effect of apatinib on apoptosis. The cell mitochondrial membrane potential changes were detected by JC-1. Then the expression changes of vascular endothelial growth factor receptor 2 (VEGFR2) pathway-related proteins were examined by Western blot.Results:Apatinib had proliferation inhibitory effects on both MV4-11 and MOLM-13 cells, and the half-maximal inhibitory concentration (IC 50) at 48 hours was (2.23±0.42) μmol/L and (4.08±2.62) μmol/L, respectively. After exposure to apatinib with increasing concentrations (10, 20, 30, and 40 μmol/L) for 48 h hours, the percentage of apoptotic cells was significantly increased in MV4-11 cells [(81.95±1.15)%, (88.80±0.23)%, (97.46±0.49)%, and (99.29±0.05)%] and MOLM13 cells [(47.30±0.87)%, (67.00±3.71)%, (82.60±2.89)%, and (98.06±5.34)%] in a dose-dependent manner, and the differences were statistically significant ( F = 6 915.0, P < 0.01; F = 5 385.0, P < 0.01). Detection of mitochondrial membrane potential by JC-1 method showed that after MV4-11 and MOLM-13 cells were treated by 10, 20, 30, and 40 μmol/L apatinib for 24 hours, the JC-1 aggregate/monomer mean fluorescence intensity (MFI) ratios were 0.45±0.06, 0.19±0.07, 0.12±0.03, 0.09±0.01, and 0.84±0.05, 0.66±0.13, 0.35±0.11, 0.27±0.02, which were different from the control group (0.67±0.15 and 0.97±0.42), and the differences were statistically significant ( F = 372.3, P < 0.05; F = 276.4, P < 0.05). Western blot was performed to detect different concentration of apatinib (2.5, 5.0 and 10.0 μmol/L) on the MV4-11 cells for 24 hours, the results showed that apatinib could down-regulate the phosphorylation of VEGFR2, Src and Stat3 in a dose-dependent manner. Conclusions:Apatinib can inhibit cell proliferation and induce apoptosis in AML with FLT3-ITD mutation. The possible mechanism is related to the down-regulation of phosphorylation of VEGFR2 and its downstream targets Src and Stat3.