Background Corneal reinnervation of subbasal nerve plexus have been found after small incision lenticule extraction(SMILE).However,there were few reports about corneal reinnervation mode and dynamic changes at the corneal cap in different time points postoperation.Objective The aim of this study was to evaluate the corneal reinnervation at the cap margin after SMILE.Methods The clinical data of 32 myopic eyes of 16 patients who received SMILE incision lenticule extraction surgery in Affiliated Eye Hospital of Shandong Medical College from April 2014 to April 2015 were retrospectively analyzed.The following-up was carried in 1 week,1 month,3 months and 6 months after surgery,and in-vivo confocal microscopy was used to acquire images of the central corneal subbasal nerve plexus before and after surgery,with the scan range of 0.4 mm×0.4 mm,from which nerve density and nerve tortuosity were evaluated using Image-Pro Plus imaging analysis software.The overall length of nerve fibers at the central cornea was measured to assess the subbasal nerve density,and the repair of nerve fibers at cap was observed.Results The corneal subbasal nerve density was (19 687.45 ± 1 147.59),(10 500.46 ± 1 056.22),(12 833.40-± 1 047.98),(13 564.04-± 1 173.01) and (14 661.35-±941.92) μm/mm2 in preoperation and postoperative 1 week,1 month,3 months,6 months,respectively,with a significant difference among different time points (F =319.44,P=0.000),and the corneal subbasal nerve densitis in postoperative time points were significantly reduced in comparison with the preoperation,and corneal subbasal nerve densities were gradually increased after surgery as the extending of time,showing significant differences between different time points (all at P＜ 0.01).Abundant corneal subbasal nerve fibers were seen with the near normal morphology after surgery.However,the fragments and disaggregation of nerve fibers were found at the incision of cap margins,and some nerve fibers extended toward the inner cap at the non-incision of cap margins 1 week after surgery.In 3-6 months after SMILE,the continuous extension of nerve fibers was exhibited under the confocal microscope.Conclusions Six months after surgery,less invasive SMILE technique can remain the superficial corneal nerve fibers.Corneal innervation shows a radiate mode from peripheral cornea outside toward inside of the cap.The subbasal never fiber density is gradually increased with the extending of time after SMILE.

Objective To improve the diagnosis and treatment of infective endocarditis(IE)by exploring its causes,pathogenic microorganism and clinicsI characteristics.Methods The clinical data of 120 IE patients treated in Peking Union Medical College Hospital from October 1997 to September 2007 were analyzed retrospectively.Results Of the 120 consecutive cascs diagnosed as IE according to the Duke's new criteria,79 were male and 41 female with a average age of(43.2±16.7)years old.Twelve cases were prosthetic valve endocarditis(PVE)and 108 cases native valve endocarditis(NVE)and there were no previously known heart diseases in 29 of the cases.Seventy-nine of the 108(73.1%)NVE patients had basic cardiac abnormalifies before IE diagnosis,such as congenital cardiovascular disease(30 cases),idiopathic mitral valve prolapse(23 cases)and rheumatic heart disease(11 cases).Fever(100.0%),anemia(54.2%)and embolism(48.3%)were the most common clinical manifestations in the IE development.Of the 83 patients who had a positive blood culture result,Streptococcus vividaus(51.8%)was the most common isolated microorganism.Conclusions Congenital cardiovascular diseases and idiopathic mitral valve prolapse are the two most commonly heart diseases in IE.Blood culture and echocardiogram should always be done to exclude IE,especially presenting with fever of unknown reasons.

Objective To prepare VP1 protein vaccine of Coxsackievirus A16(CA16) and evalu-ate immunngenicity the subunit vaccines of Coxsackievirus (VP1), and to establish foundation for studying CA16 vaccine. Methods CA16 VP1 was amplified by RT-PCR and cloned into pFastBac HT A plasmid, recombinated with Bacmid DNA by transposition reaction and then transfected Sf9 cell, mixed with adjuvant AI(OH)_3. After immunization BALB/c mice, evaluating immune effectiveness after booster injections 2 weeks. Results The expressed protein was analyzed by SDS-PAGE and Western blot, mice immunized with CA16 (VP1) both induced specific IgG antibody and neutralization antibody. The best immunization antigen was 20 μg, IgG antibody was 1: 1600, neutralization antibody was 1:250, typical Th1/Th2 immune response was determined by lymphocyte proliferation assay and cytokine analysis. Conclusion The CA16 VP1 gene was cloned successfully and expressed in Sf9 insect cells, CA16 VP1 protein vaccine induced both humoral and cellular immune response, to lay solid foundation for further study on CA16 vaccine.

Objective To explore the value of spectral imaging technique in dual-energy CT in differential diagnosis of the metastatic lymph nodes in thyroid carcinoma,squamous cell carcinoma metastatic lymph nodes and lymphoma in the neck.Methods In 30 patients with pathologically confirmed with a total of 79 cervical lymph nodes enlargement which were using dual energy scan .Then observed the change trend of the spectrum curve and comparison the three kinds of lymph node energy spectrum curve’s slope.Results In the 79 lymph nodes,the metastatic lymph nodes in thyroid carcinoma were twenty-three,squamous cell carcinoma metastatic lymph nodes were twenty-four and lymphoma were thirty-two.From 60 to 180 keV,with the increase of keV values,the three kinds of malignant lymph nodes of the corresponding CT value decreasing and the higher the keV value,the CT value decrease magnitude was small,and the spectrum curve was〞drop type〞.The slope spectrum curve of the metastatic lymph nodes of thyroid carcinoma in arterial phase and parenchymal phase were maximum,which were 1.23±0.41 and 0.85±0.33,respectively.The slope spectrum curve of lymphoma in arterial phase and parenchymal phase were least,whcih were 0.40±0.16 and 0.47 ±0.09.The slope spectrum curve of the squamous cell carcinoma metastatic lymph nodes in arterial phase and parenchymal phase were 0.88±0.10 and 0.62±0.28.The energy spectrum curve slope of the three kinds of malignant lymph nodes have statistical significance.Conclusion The energy spectrum curve slope of arterial phase and parenchymal phase has some significance lymph node metastasis of in thyroid carcinoma,the metastatic lymph nodes and lymphoma in the neck.

Objective To set up a technical platform of reverse genetics based on the 8 plasmid.virus rescue system of cold-adapted influenza virus strain. Methods The cold-adapted, temperature sensitive, live attenuated influenza virus strain A/AnnArbor/6/60(H2N2)was chosen as the master donor virus(MDV)for rescue research,and its six internal gene fragments PB2,PB1,PA,NP,M and NS were artificially synthesized. Meanwhile, five amino acid mutations have been introduced as tags. Six fragments were ligated with modified pAD3000 for the construction of rescue plasmid. Six transcription/expression plasmids(pMDV-A-PB2,pMDV-A-PB1,pMDV-A-PA,pMDV-A-NP,pMDV-A-M,and pMDV-A-NS)were obtained, and their sequences were accurate. Results The reassorted virus named as rMDV-A contains HA and NA gene segments derived from PR8 strain along with six gene segments,PB2,PB1,PA,NP,M and NS,from MDV. The COS-1 cells were co-transfected with eight recombinant plasmids. The results showed that a cold-adapted, attenuated reassortant influenza A virus with hemagglutination activity was rescued successfullv bv"6+2" combination of MDV and PR8, and the allanotoic fluid of the injected eggs gave a posigenes of A/AA/6/60 used as backbone has provided experimental materials for further research on the gene function and novel vaccine candidate of cold-adapted, attenuated human influenza virus.

Six gene segments,PB1,PB2,PA,NP,M and NS,were fully synthesized which derived from the master donor virus (MDV),cold-adapted(ca),temperature sensitive(ts),live attenuated influenza virus strain A/Ann Arbor/6/60(MDV-A).Meanwhile,five amino acid substitutions (PB1-391E,58lG,661T,PB2-265S,NP-34G) were artificially altered by human intervention.HA and NA fragments derived from the 2006-2007 circulating strain A/New Caledonia/20/99 (H1N1).Eight fragments were ligated with modified pAD3000 for rescue plasmid construction.Eiight transcription/expression plasmids were named as pMDV-A-PB2,pMDV-A-PB1,pMDV-A-PA,pMDV-A-NP,pMDV-A-M,pMDV-A-NS,pMDV-A-HA,pMDV-A-NA,respectively.The COS-l cells were co-transfected with eight plasmids representing 6 internal viral backbone of the strain A/AA/6/60 and two plasmids containing the CDNA of the HA and NA segments of the strain A/New Caledonia/20/99 (H1N1),the results showed that cold-adapted,attenuated reassortant influenza A virus Was rescued successfully.Titers of a reassorted influenza A virus in embryonated chicken eggs mnged from 1:29to l:210.The rescue system of six intemal genes used as backbone opens the way for further research on gene function and neotype vaccine candidate of cold-adapted,live attenuated human influenza virus.

Six gene segments, PB1, PB2,PA, NP, M and NS, were fully synthesized which derived from the master donor virus(MDV), cold-adapted(ca),temperature sensitive(ts), live attenuated influenza virus strain A/Ann Arbor/6/60(MDV-A). Meanwhile, five amino acid substitutions (PB1-391E, 581G, 661T, PB2-265S, NP-34G) were artificially altered by human intervention. HA and NA fragments derived from the 2006～2007 circulating strain A/New Caledonia/20/99 (H1N1). Eight fragments were ligated with modified pAD3000 for rescue plasmid construction. Eight transcription/expression plasmids were named as pMDV-A-PB2, pMDV-A-PB1, pMDV-A-PA, pMDV-A-NP, pMDV-A-M, pMDV-A-NS, pMDV-A-HA, pMDV-A-NA, respectively. The COS-1 cells were co-transfected with eight plasmids representing 6 internal viral backbone of the strain A/AA/6/60 and two plasmids containing the cDNA of the HA and NA segments of the strain A/New Caledonia/20/99 (H1N1), the results showed that cold-adapted, attenuated reassortant influenza A virus was rescued successfully. Titers of a reassorted influenza A virus in embryonated chicken eggs ranged from 1∶29 to 1∶210. The rescue system of six internal genes used as backbone opens the way for further research on gene function and neotype vaccine candidate of cold-adapted, live attenuated human influenza virus.

A cold-adapted (ca), temperature sensitive (ts), live attenuated influenza B virus strain B/Ann Arbor/1/66 was chosen for influenza virus rescue research, in which six internal gene segments, PB1, PB2, PA, NP, M, NS, were fully synthesized and nine amino acid substitutions were artificially alter by human intervention. The resultant B/Ann Arbor/1/66 plasmids were named as pAB121-PB1, pAB122-PB2, pAB123-PA, pAB124-HA, pAB125-NP, pAB126-NA, pAB127-M and pAB128-NS, respectively. A recombinant influenza A virus was previously generated entirely from cloned cDNA. An infectious recombinant influenza B virus was generated here, and designated as rMDV-B, by plasmid-based reverse genetics. The rMDV-B virus contained HA and NA genes from an epidemic influenza B vires strain B/Malaysia/2506/2004 in the background of internal genes derived from influenza B virus strain B/Ann Arbor/1/66. HA titer of rMDV-B in MDCK cells and embryonated chicken eggs ranged from 1 : 64 to 1 : 512. The results may allow an effective live influenza B vaccine to be produced from a single master strain, providing a model for the design of future live human influenza vaccines.

Objective:To the investigate the predictive value of preoperative systemic inflammatory response index (SIRI) for postoperative clinical outcome in patients with spontaneous intracerebral hemorrhage (ICH) in basal ganglia region.Methods:Patients with ICH in basal ganglia region underwent surgical treatment at the Department of Neurosurgery, the Affiliated Hospital of Qingdao University from January 2015 to December 2021 were retrospectively included. At 3 months after surgery, the modified Rankin Scale was used to evaluate the clinical outcome, with a score of 0-2 defined as good outcome and >2 defined as poor outcome. Multivariate logistic regression analysis was used to identify the independent correlation between SIRI and poor outcome in patients with ICH. Receiver operating characteristic (ROC) curve analysis was used to identify the predictive value of SIRI. Results:A total of 258 patients with ICH in basal ganglia region underwent surgical treatment were enrolled, including 176 males (68.22%), aged 57.00 years (interquartile range, 49.00-65.25 years); median hematoma volume was 50.00 ml (interquartile range, 40.00-70.00 ml), and median SIRI was 4.12 (interquartile range, 2.28-7.30); 143 patients (55.43%) had poor outcome. Multivariate logistic regression analysis showed that older age (odds ratio [ OR] 1.070, 95% confidence interval [ CI] 1.030-1.111; P<0.001), lower Glasgow Coma Scale score ( OR 0.669, 95% CI 0.575-0.779; P<0.001), higher platelet count ( OR 1.010, 95% CI 1.003-1.017; P=0.004), and higher SIRI ( OR 1.434, 95% CI 1.255-1.638; P<0.001) were the independent predictors of poor outcome. ROC curve analysis showed that the area under the curve for predicting poor outcome by SIRI was 0.791 (95% CI 0.737-0.845; P<0.001), with an optimal cutoff value of 4.53. The predictive sensitivity and specificity were 67.8% and 81.7%, respectively. Conclusion:Preoperative SIRI can effectively predict the clinical outcome of patients with ICH in basal ganglia area at 3 months after surgery, and SIRI >4.53 indicates poor outcome.

Objective:To investigate the effect and of piperine(PIP)on nerve injury in rats with cerebral infarction.Meth-ods:The rat model of cerebral infarction was prepared by modified thread embolism method.The rats were divided into Sham group,Model group,piperine group(PIP group,20 mg/kg piperine),piperine+PKA inhibitor group(PIP+H-89 group,20 mg/kg PIP+5 mg/kg H-89).The rats were scored for neurological impairment,the volume of cerebral infarction,neuronal damage and neuronal apoptosis were observed,and the levels of cAMP,IL-1β and IL-6 in hippocampus and the protein expressions of GFAP,NSE,AC6,PKA,p-CREB,CREB and BDNF were detected.Results:Compared with Sham group,Model group had large cerebral infarction,cerebral in-farction volume,the neurological deficit score,neuronal apoptosis rate,the levels of IL-6 and IL-1β in hippocampus,and number of GFAP positive cells,the positive expression rates of NSE protein increased,the number of Nissl bodies,and the expressions of cAMP,AC6,PKA,p-CREB and BDNF proteins in hippocampus decreased(P<0.05);compared with Model group,cerebral infarc-tion volume,the neurological deficit score,neuronal apoptosis rate,the levels of IL-6 and IL-1β in hippocampus,and number of GFAP positive cells,the positive expression rates of NSE protein in the PIP group decreased,the number of Nissl bodies,and the ex-pressions of cAMP,AC6,PKA,p-CREB and BDNF proteins in hippocampus increased(P<0.05);the effect of piperine in improving nerve injury in rats with cerebral infarction was able to be reversed by H-89.Conclusion:PIP can inhibit the apoptosis of hippocampal neurons,and improve the neural injury induced by cerebral infarction by activating AC/PKA/CREB signal pathway.