ObjectiveTo construct self-amplifying RNA(saRNA)vaccine of severe acute respiratory syndrome coronavirus2(SARS-CoV-2)Delta mutant strain(B.1.617.2)based on Coxsackievirus-A5(CV-A5)replicon and evaluate its immunogenicity.MethodsThe recombinant plasmids pDelta-S10,pDelta-S5 and pDelta-S1(10,5 and 1 amino acid residues at the upstream of S-VP1/2A cleavage site of the fusion polyprotein respectively)were constructed by In-fusion cloning of the plasmids containing the full-length genome sequence of CV-A5 and substituting the S protein gene of SARS-CoV-2 Delta mutant for the P1 structural protein gene of CV-A5 with different lengths.Three RNA molecules,Delta-S10,Delta-S5 and Delta-S1,were obtained by in vitro transcription of linearized recombinant plasmids and transfected into HEK-293T cells respectively,which were analyzed for the expression of S protein by Western blot.The RNA molecule with the highest expression of S protein was screened out and detected for the self-amplification in HEK-293T cells by qPCR.BALB/c mice(female,6 ～ 8 weeks old and five for each group)were immunized i.m.with two doses(0.5 and 2.5 μg)of the screened Delta-S packaged with lipid nanoparticles for once on day 1 and day 14 seperately.Blood samples were collected on days 14and 28,detected for serum binding antibody titers by ELISA,and detected for neutralizing antibody titers by micro neutralization method.The spleens were harvested on day 42 and detected for the level of IFNγ secreted by mouse spleen cells by enzyme linked enzyme linked immunospot assay(ELISPOT).ResultsThe recombinant RNA molecule Delta-S10showed the highest expression of S protein and self-amplified in HEK-293T cells,which of both high and low doses induced specific binding antibody against SARS-CoV-2 Delta S1 protein in mice with obvious dose effect and enhanced immune effect;The high dose of Delta-S10 induced neutralizing antibodies and cellular immune responses in mice.ConclusionThe SARS-CoV-2 Delta mutant(B.1.617.2)saRNA vaccine Delta-S10 based on CV-A5 replicon was successfully constructed,which induced humoral and cellular immune responses in mice,laying a foundation of the further study of the construction of SARS-CoV-2 saRNA vaccine by enterovirus replication elements.

Objective Recent researches indicate that Eph/ephrin signaling pathway is possibly related to adult neurogenesis after cerebral injury..With brief introduction of their structures and interactions,the review focus on their possible mechanism in adult neurogenesis.Methods The literatures between 2010 and 2015 were retrieved following online databases:PUBMED,ScienceDirect,Wanfang and CNKI database.The key words used in the reasearch were Eph,ephrin,cerebral injury,adult neurogenesisand so on.Results Totally 42 related articles were enrolled.And these papers discribed how researchers performed their experiments and further explained Eph/ephrin 's vital roles in adult neurogenesis.Conclusion Eph/ephrin signaling can influence adult neurogenesis after cerebral injury.positively or negatively.And Akt may be a downstream signaling molecule of Eph/ephrin that change the progress of adult neurogenesis.However,the detailed mechanisms remain to be further study.

Objective To investigate the predictive value of neutrophil and lymphocyte ratios (NLR)for the prognosis in patients with acute cerebral infarction. Methods From January 2014 to December 2015,307 consecutive patients with acute cerebral infarction admitted to the Department of Neurology,the Fifth Affiliated Hospital of Zhengzhou University were enrolled retrospectively,including 80 females and 227 males. They were divided into ether a good prognosis group (n = 195)or a poor prognosis group (n = 112)according to the scoring criteria of the modified Rankin scale (mRS). The age,gender, past medical history,National Institutes of Health stroke scale (NIHSS)score were documented on admission. The NLR values were calculated according to the neutrophil and lymphocyte counts on admission. Logistic regression analysis was used to analyze the influencing factors of poor prognosis of acute cerebral infarction. The receiver operating characteristic curve (ROC)was used to evaluate the predictive effect of the NLR level on patients with acute cerebral infarction on admission. Results (1)Compared with the good prognosis group,the age,incidence of recurrent cerebral infarction,NIHSS score on admission, NLR levels on admission in the poor prognosis group were higher. There were significant differences between groups (69 ± 12 years vs. 62 ± 14 years,25. 0% [28 / 112]vs. 14. 4% [28 / 195],5. 00 [3. 00, 9. 00]vs. 3. 00 [1. 75,5. 00],and 3. 66 [2. 62,7. 91]vs. 2. 47 [1. 94,3. 40];all P < 0. 05). There were no significant differences in other baseline data and clinical characteristics between the groups (all P >0. 05). (2)Multivariate logistic regression analysis showed that the increase of the age,NLR level on admission,and increased NIHSS score on admission,were independent risk factor for poor prognosis (OR 1. 030,1. 148,and 1. 427,respectively,95% CI were 1. 007 -1. 053,1. 059 -1. 246,and 1. 247 -1. 634, respectively;all P < 0. 05). (3)The diagnostic cut-off value of the NLR level on admission for the poor prognosis in patients with acute cerebral infarction was 2. 84. Its sensitivity was 69. 6% and specificity was 64. 6% . Conclusion The increase of the NLR level on admission had certain reference function on the poor prognosis in patients with acute ischemic stroke.

Objective To evaluate the diagnostic accuracy of transcranial doppler(TCD)as an additional screening tool for intracranial arterial steno-occlusive disease in patients with acute ischaemic stroke(AIS)by comparing with the computed tomography angiography(CTA). Methods Two hundred and twenty-four consecutive patients who were hospitalized within 7 days of the onset of symptoms and fulfilled the criteria for the clinical di-agnosis of AIS between May 2013 and May 2014 were enrolled for the study. Among the 224 patients,there were 118 males and 106 females with an average age of 61.2±12.6 years. High-resolution brain CTA was performed after completion of TCD. Results There was slight distinction of the diag-nostic accuracy of TCD for different arteries. TCD demonstrated the most accurate diagnosis for carotid artery vascular system,with a sensitivity of 96.19%and specificity of 98.60%. For the vertebrobasilar vascular system,TCD diagnosis showed a sensitivity of 71.11%and specificity of 94.31%. TCD findings were complementary to the CTA results(real-time embolization,collateral flow patterns and steal phenomenon). Conclusion TCD shows a high diagnostic accuracy comparing to CTA in evaluating intracranial and extracranial arterial stenosis in patients with AIS,especially for the carotid artery vascular system analysis within a short time interval. TCD can also provide additional real-time dynamic findings complementary to the information provided by CTA.

Objective To identify the types of pulmonary complications after pancreaticoduodenectomy,and to discuss the prevention and management of these complications.Methods Clinical data of 165 cases of pancreaticoduodenectomy in our hospital were retrospectively analyzed.Pulmonary complications were identified,therapeutic effects were observed.Results The incidence rate of pulmonary complications was 19.4％ ( 32/165),case-fatality rate was 6.25％ (2/32),two patients died from pneumonia,respiratory failure and ARDS.Complications mainly included pneumonia 13.9％ (23/165),pleural effusion 4.2％ (7/165),atelectasis 3.6％ (6/165),pneumothorax 1.8％ ( 3/165 ),respiratory failure 2.4％ (4/165) and ARDS 1.2％ (2/165).Conclusion Pulmonary complications after pancreaticoduodenectomy are not rare,especially for pulmonary infection and most are hospital acquired pneumonia.To understand rules and particularity of respiratory physiopathological changes after pancreaticoduodenectomy is very important for patients to safely pass over the perioperative period.

Objective To investigate the clinical characteristics, diagnosis, treatment and prognosis of pancreatic gastrointestinal stromal tumor (GIST). Methods We reported a case and reviewed the medical literature on pancreatic malignant GIST. We searched the Pubmed and main domestic database. The clinical data of the reported cases were studied, and their predictive factors for postoperative recurrence and metastasis were analyzed. Results Between January 1980 and July 2010, 16 cases of pancreatic GIST were reported. There were 7 males and 9 females, with a median age pf 56.5 (31-72)years. The clinical symptoms were nonspecific. The main presentation was upper abdominal pain or discomfort. A preoperative diagnosis was suspected on radiological examination. The tumor mainly appeared as a well-defined solid-cystic mass. Irregular enhancement appeared in the circumferential and solid portion of the tumor on enhanced CT scan sequences. The pancreatic and biliary ducts were rarely dilated. Endoscopic ultrasound guided fine needle aspiration cytology (EUS-FNA) was helpful in preoperative diagnosis. Of the 15 surgical patients, 14 underwent complete resection, while the remaining received cyst-jejunostomy. A correct diagnosis was made on histopathology and immunohisto-chemistry. On a mean follow up of 21 months (range, 1-60) in 14 patients, all patients were alive.Recurrence or metastasis occurred in 4 patients with tumors of high malignant potential. On univariate analysis, the only significant predictor for adverse outcome was mitoses≥10/50 HPF. Conclusions Pancreatic GIST is a rare tumor of relatively low malignant potential. It has a better prognosis than ductal adenocarcinoma. It is important to arrive at a correct diagnosis and treat the tumor with radical resection. Aggressive surgical resection is potentially curative. Imatinib is recommended in the treatment of patients with tumors with high malignant potential.

Objective To investigate the dynamic changes of microglial polarization at the perihe-matoma area and provide timepoint evidence for interventing microglial polarization as well as studying the polarization mechanism after intracerebral hemorrhage ( ICH ) . Methods Healthy male Sprague Dawley (SD) rats were randomly divided into sham group,ICH-4 h,1 d,3 d,7 d and 14 d groups with 6 in each group. The rats in ICH groups were injected collagenase VII-s into the caudate nucleus to establish the in-tracerebral hematoma model and rats in sham operated group were treated with the same amount of saline. The brains were taken at 4 h,1 d,3 d,7 d,14 d in the ICH group,1 d in sham group. Microglia typeⅠ( M1, CD11b++CD86+) and microglia typeⅡ( M2,CD11b++Arg-1+) were examined by immunofluorescence and the number of M1 and M2 around hematoma were analyzed. Results ( 1) The M1 and M2 were both ob-served at 4 h after ICH and a small quantity of branches were still presented on M1. ( 2) M1 took the main position in acute stage (1~3 d),early subacute stage(3~7 d) and chronic stage (>14 d) after ICH.The number of M2 was elevated transiently in superacute (<24 h) and late subacute stage (7 d).The number of M2 (31.40±1.69) was more than M1 (21.43±1.81) at 4 h after ICH ( t=- 4.085, P=0.002),and the number of M2 (116.25±5.06) significantly exceeded M1 (85.75±7.32) again on day 7 ( t=-0.690, P=0.001). Conclusion M1 is in a dominant position in acute,early subacute and chronic stages after ICH;M2 is dominant in superacute and late subacute stages. Investigating the mechanism of M2 formation at acute period ( such as 4 h) or late subacute stage ( such as 7 d) ,and inhibiting M1 formation in the early subacute stage ( 1~3 d) have important significance for clinical treatment of ICH.

Objective To evaluate the efficacy of preoperative regional intra-arterial chemotherapy (RIAC) in the treatment of resectable pancreatic head carcinoma. Methods The clinical data of 50 patients with resectable pancreatic head carcinoma who had been admitted to the Research Institute of Pancreatic Diseases of Fudan University from December 2006 to July 2007 were retrospectively analyzed. Patients were randomly divided into2 groups (n =25 in each group): patients in group A were treated with preoperative RIAC followed by regional pancreaticoduodenectomy, and patients in group B were treated with surgical procedure routinely. The lymphatic metastases in the 50 specimens of pancreatic head carcinoma were detected by histological examination with hematoxylin and eosin (HE) staining, and lymphatic micrometastases were detected by immunohistochemical method with staining of cytokeratin AE1/AE3 in 10 specimens with negative HE staining of the lymph nodes in each group. Results There was no significant difference in the incidence of complications, the length of hospital stay and the 1-, 2-year survival rates between the 2 groups (χ2 = 0.12, 2.88, P > 0.05). The incidence of positive lymph node metastasis in group A was 7.1% (52/734), which was significantly higher than 22.1% (118/532) in group B (χ2 = 60.01, P < 0.05). The incidence of lymphatic micrometastasis was 9.4% (30/319) in group A, and 9.1% (23/252) in group B, with no statistical difference between the 2 groups (χ2= 0.01, P > 0.05). Conclusions Preoperative RIAC is helpful in improving the prognosis of patients with resectable pancreatic head carcinoma by reducing the incidence of lymphatic metastasis and decreasing tumor stage.

Objective To explore the mechanisln of action of Jianpiyishen Decotion on renal fibrosis and provide experimental evidence for chnieal application.Methods The Wistar male rats were randomly divided into four groups.They were normal control group(N group),model group(M group),low dose treatment group(L group)and high dose treatment group(H group).AⅡanimal models were made of CRF with subtotal renal ablation except the N group.Interference began at one week after operation.After being interfered for 8 weeks,blood serunl and nephridial tissues were taken out.Serum urea nitrogen(BUN)and creatinine(Scr)and fibronectin(FN)were detected.Results In N group。There were light expression in renal tubniointerstitial substance,cellula epithelialis basal meulbnule and Vascular smooth muscle.In M group,FN was strong expressed in renal tubulointerstitial substance and renal tubde,and also in glomerular mesangium.There was significant difference between N and M group(P

A 33-year-old male patient complained of presenting goiter on the low back area for 2 months. Pathological examinations of resected goiter suggested non-Hodgkin lymphoma and showed that T cells, immunoblasts, and hemogram were roughly normal, and 2% sarcoma cells could be found in bone marrow. Stage Ⅳ T-cell non-Hodgkin's lymphoma was diagnosed. Following 4 months of chemotherapy using CHOP protocol (cyclophosphamide, adriamycin, vincristine, and prednisone included), the patient underwent bone marrow activation and autologous peripheral blood stem cell transplantation in combination with recombinant interleukin-2 application in April 1998. The preprocessing was performed under MACC protocol (L-sarcolysinum, cytarabine, cyclophosphamide, and Iomustine included). Ten days after autologous stem cell transplantation, neutrophil concentration was > 0.5×109/L and sixteen days after transplantation, blood platelet concentration was > 50×109/L. Six days after transplantation, the patient exhibited fever, and E. Coli infection was confirmed through blood culture. After antibiotic treatment, body temperature recovered to normal, and fever disappeared. The patient had been followed-up for 10 years and 10 months. During the follow-up period, he lived a normal life and work.