Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

Objective To evaluate the value of creatine kinase MB(CK-MB) and cardiac troponin I(cTnI)to earlier diagnosis on myocardial injury in newborn infants with asphyxial.Methods Dynamic variation of serum CK-MB and cTnI levels were measured at birth 1,5 and 10 days,respectively,in 40 asphyxia newborn infants and 20 control neonates.Results Serum CK-MB and cTnI levels of asphyxia neonates were significantly higher than those in control group(P0.05).Conclusion The determination of CK-MB and cTnI levels can help the prediction of myocardial injury after asphyxia.

Objective To detect the expression pattern of microRNA in A549 cells treated by lipopolysaccharide, study the expression of miRNA-1247-3P in A549 cells under LPS treatment and explore the possible mechanism of miRNA-1247-3P in A549 cells under LPS treatment. Methods A549 cells were divided into experimental and control groups. Immunocytochemical method and RT-PCR were used to detect the changes of SP-A and SP-C. The expression of miRNAs were detected using miRNAs array in different groups. The key miR-1247-3P was collected to detect the changes of miR-1247-3P in all groups using quantitative real-time polymerase chain reaction. Results Compared with control group, the expressions of SP-A and SP-C were significantly decreased in the experimental groups (P < 0.05). MiRNA array showed that 31 miRNAs were up-regulated and 3 miRNAs were down-regulated. Compared with control group, the expression of miR-1247-3P was significantly increased in the experimental groups (P < 0.05). Conclusion The increased expression of miR-1247-3P may play an important role in the pathogenesis of ARDS.

Objective Investigate the reasonable surgical approach combined with frozen sections pathological features of papillary thyroid microcarcinoma.Methods Clinical data of 96 cases papillary thyroid microcarcinoma nearly 5 years of follow-up in our clinic referral was retrospectively analyzed.Metastasis and recurrence rate were compared between group of ipsilateral lobe plus isthmus resection (unilateral group) and group of ipsilateral lobe plus isthmus resection and contralateral lobe subtotal (bilateral group).Metastasis and recurrence rate were compared between group of central lymph node dissection (dissection group) and non-dissection group (non-dissection group),and the injury rate of the recurrent laryngeal nerve was also compared between the two groups.Results The diagnosis of cancer by intraoperative frozen pathology were 53 cases (55％).Whether in high or low risk populations,the metastasis and recurrence between unilateral and bilateral groups showed no significant difference (P ＞ 0.05).Whether in high or low risk populations,the metastasis and recurrence between dissection group and non-dissection groups showed no significant difference (P ＞0.05).The temporary injury rate of recurrent laryngeal nerve in dissection group were higher than thatin non-dissection group both in high-risk populations (P =0.040,P ＜ 0.05) and low risk populations(P =0.037,P ＜ 0.05).Conclusions Intraoperative frozen pathological diagnosis of papillary thyroid microcarcinoma is difficult.The reasonable surgical approach for the first time is ipsilateral lobe plus isthmus resection.Preventive cervical dissections operation should not be carried out if the exploratory of lymph node showed no metastasis.

Objective:To evaluate the prediction value of preoperative Fibrinogen(FIB) in combination with neutrophil-lymphocyte ratio(NLR)for the prognosis of operable pancreatic cancer patients.Methods:The clinical data of 124 patients who underwent radical resection for pancreatic cancer in our hospital from Jan 2010 to Dec 2018 were retrospectively analyzed.The patients were divided into three groups according the high and low NLR, FIB value which defined by the receiver operating characteristic curve (ROC). The clinicopathological data and overall median survival time were compared between the three groups.Results:Univarate analysis showed that age, tumor stage, NLR, FIB and F-NLR score were associated with the prognosis( P<0.05), while multivariate analysis showed that high F-NLR score was the independent prognostic factor. The median survival time of patients with F-NLR scores 0, 1 and 2 group was 30.6, 20.3 and 13.9 months( P<0.05). The prognosis of high F-NLR score was significantly worse than that of low F-NLR score( P<0.05). Conclusions:A high preoperative F-NLR score was a promising predictor for the prognosis of pancreatic cancer patients after radical resection.

OBJECTIVE: To investigate the expression of monocyte chemotactic protein-1 (MCP-1) and its receptor CC chemokine receptor-2 (CCR-2) in coronary atherosclerosis plaques between sidden coronary death (SCD) and non-SCD. Methods The expression levels of MCP-1 and CCR-2 in SCD group, coronary atherosclerosis group (non-SCD), control group (normal coronary artery) were detected by immunohistochemistry. RESULTS: Positive rates of MCP-1 among the three groups were 78%, 47%, and 0%, respectively, with significant expressing differences between each two groups (P<0.05). Positive rates of CCR-2 among three groups were 72%, 47%, and 0%, respectively, with significant expressing differences between the SCD group and coronary atherosclerosis group as well as between the SCD group and control group (P<0.05), but with no significant expressing difference between coronary atherosclerosis group and control group (P>0.05). CONCLUSION Overexpression of MCP-1 and CCR-2 in coronary atherosclerotic plaques is closely correlated with SCD.
Chemokine CCL2/metabolism* ; Coronary Artery Disease/pathology* ; Death, Sudden, Cardiac/pathology* ; Humans ; Immunohistochemistry ; Receptors, CCR2/metabolism*

Objective To establish and evaluate 3 kinds of minimally-invasive valve implantation model in vivo.Methods A novel tissue engineered heart valve(TEHV) manufactured by branched polyethylene glycol cross-linked acellular porcine valve and a minimally-invasive valve implantation system according to the design of Corevalve revalving system were adopted.After anesthesia,18 adult male goats were randomly divided into 3 groups: the ulrasound-directed orthotropic group (group A,n =6),angiography-directed orthotropic group (group B,n =6) and direct-released heterotopic group (group C,n =6),and all received minimally-invasive valve implantation orthotropically or heterotopically.4 weeks later,the valvular function was evaluated by CTA and/or echocardiography.Results All 3 kinds of caprine model were successfully constructed.The operation success rate of each group was A: 66.7％,B: 50.0％ and C: 100.0％,respectively(multiple x2 analysis,group A and B P ＞0.05; group A and C,group B and C,P ＜0.05).The operation-time of each group was A: (79 ± 18) min,B:(61 ±23) min,C: (45 ± 15) min(one-way ANOVA,P ＜0.05).The survival rate at4 weeks was A: 100％,B: 100％ and C: 83.3％ (multiple x2 analysis,P ＞ 0.05).Echocardiography and CTA proved the short-term function of implanted TEHV was satisfactory.Conclusion All 3 kinds of caprine valve implantation model can be established without cardiopulmonary bypass and blood transfusion.The devices and equipments required in group A is relatively simple,but the procedure cost longer time for it is hard to determine the right position by ultrasound.The application of angiography made the positioning much easier in group B while the procedure had to be performed in specific operation room with angiographic apparatus.Group C did rely on neither special equipments nor complex operation,but the valve leaflets cannot work normally,so this model was only suitable for testing in vivo characteristics such as biocompatiblities.

To observe the effects of ultrasound-guided stellate ganglion block (SGB) on cerebral oxygen metabolism and postoperative cognitive dysfunction (POCD) of elderly patients, we collected 80 elderly patients undergoing selective coronary artery bypass graft under cardiopulmonary bypass. The Mini Mental State Examination (MMSE) was applied to test the cognitive function. The SjvO2, Da-jvO2 and CEO2 were used for the analysis of the cerebral oxygen metabolism. We found that POCD was related to disequilibrium of cerebral oxygen metabolism. Ultrasound-guided SGB before surgery reduced the incidence of POCD because of the improvement of cerebral oxygen metabolism.
Aged ; Autonomic Nerve Block ; Cardiopulmonary Bypass ; Cognition Disorders ; prevention & control ; Coronary Artery Bypass ; Humans ; Neuropsychological Tests ; Oxygen Consumption ; Postoperative Complications ; Stellate Ganglion ; diagnostic imaging ; Ultrasonography

National Natural Science Foundation of China becomes a dominating competitiveness that reflects to hospitals' ability of scientific research in China.Based on application and fund availability of National Natural Science Foundation of Shanghai 6th people's hospital in recent years,this article compared data with other hospital's condition to analyze shortcoming and challenge.It also gave suggestions of selecting a subject,raising rate of support,training outstanding young researchers,enhancing interdisciplinary study,and advancing quality of academic papers.This article would give a reference to promote high-speed,harmonic and sustainable development of scientific research in our hospital.

Objective: To screen candidate antigen for therapeutic HBV vaccine with a novel adjuvant DC-Chol. Methods: BALB/c mice were injected with DC-Chol liposome and HBsAg prepared from CHO and Yeast respectively. One week later, IL-4, IL-2, IFN-?were measured by ELISA or ELISPOT. Results: The levels of IL-2, IFN-?of HBsAg from Yeast with DC-Chol liposome were 20 and 119 times higher respectively than those of HBsAg from CHO with DC-Chol liposome. ELISPOT assay showed that the counts of spot-forming cells of IL-4 and IFN-?of HBsAg from Yeast with DC-Chol liposome were 2.8 and 46.3 times higher respectively than those of HBsAg from Yeast with Al(OH)3. Conclusion: HBsAg prepared from Yeast together with DC-Chol liposome may be an appropriate candidate for therapeutic HBV vaccine .