BACKGROUND: Tissue-engineered skin plays an important role in the repair and reconstruction of large-area skin lesion. Little is known about the effects of Chinese medicine-loading tissue-engineered skin scaffold on cell adhesion and wound surface infection. OBJECTIVE: To screen the application concentration of shengji solution, which can promote tissue regeneration, using keratinocytes and fibroblasts, and to observe the effects of shengji solution-gelatin-chitosan drug-loading skin scaffold on cellular adhesion and biocompatibility. DESIGN, TIME AND SETTING: Taking keratinocytes and fibroblasts as subjects, the present randomized and controlled experiment was performed at the Laboratory of Cell Engineering, Orthopedic Institute, Tianjin Hospital between February and August 2005. MATERIALS: One healthy big-ear rabbit was included for harvesting skin seed cells. Shengji solution, Danggui (Radix Angelicee Sinensis) and Shengdi (rehmannia dride rhizome) extract (self-extracted, 1.5 g/mL), gelatin-chitosan scaffold and shengji solution-gelatin-chitosan scaffold were provided by Tianjin University, China. METHODS: Passage 3 keratinocytes and fibroblasts were harvested by dispase Ⅱ- trypsase- ethylenediamine tetraacetic acid(EDTA) digestion. The prepared cells were divided into 5 groups: control, shengji solution, Danggui, Shengdi, and epidermal growth factor (EGF). Common culture solution containing 0.1 volume fraction of fetal bovine serum, shengjisolution (5, 6, 8, and 12 g/L), Danggui extract (8 g/L), Shengdi extract (8 g/L), and EGF culture medium (10 μ g/L) were used in corresponding groups for cell culture. At 1, 3, 5, and 7 days, cellular proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. Keratinocytes and fibroblasts were cultured in different concentrations of shengji solution (50, 60, 80, and 120 g/L) and grouped according to different concentrations. At 7 and 14 days, cellular adhesion was observed by semi-quantitative method. At 4, 7, and 14 days, keratinocytes cultured by 60 and 80 g/L shengji solution were harvested for hematoxylin-eosin staining and subsequent scanning electron microscope observation. MAIN OUTCOME MEASURES: Effects of shengji solution on skin seed cell proliferation; effects of drug-loading scaffold on cellular adhesion; and cell-carrier biocompatibility. RESULTS: MIF detection results demonstrated that cells significantly proliferated after treatment of 5, 6, and 8 g/L shengji solution compared to the control group (P < 0.05). Obvious proliferation of passage 3 keratinocytes and fibroblasts was found on 60 and 80 g/L drug-loading scaffold. Keratinocytes on the drug-loading scaffold exhibited good cellular morphology and closely adhered to the scaffold. Shengji solution had no apparent toxic effects on cells. CONCLUSION: Shengji solution (60 and 80 g/L)-gelatin-chitosan scaffold can effectively promote the adhesion and proliferation of keratinocytes and fibroblasts.

Objective To evaluate the value of procalcitonin(PCT) and endotoxin(ET) in early diagnosis and prognosis in critically illness patients with sepsis.Methods A total of 104 cases with suspected sepsis were prospectively collected in this study from February 2015 to October 2016 in ICU.There were 31 cases of systemic inflammatory response syndrome (SIRS group),30 cases of sepsis 24 cases of severe sepsis and 19 cases of septic shock.Prognosis was observed in patients with sepsis discharged,57 cases were survival (survival group) while 16 cases were dead (death group).The level of PCT and ET and the clinical characteristics were recorded,The the ROC curve was applied to evaluate the discriminative power of variables.Results The levels of PCT,APACHE-Ⅱ score,ET of cases with sepsis,severe sepsis and septic shock were significantly higher than those in SIRS group(P<0.05).The PCT and APACHE-Ⅱ scores were elevated by the severities of disease (P<0.05).ROC curve analysis showed that,the sensitivity of PCT value was 82.2% and specificity was 87.5%.PCT and APACHE-Ⅱ scores were higher in the death group (P<0.05),whereas there was no significant difference on ET between the two groups (P>0.05).ROC curve showed that AUC of PCT was 0.867,which was significantly higher than that for APACHE-Ⅱ score (AUC=0.762) (P>0.05).Conclusion Compared with APACHE-Ⅱ score and ET,PCT can help clinicians to early diagnose and treat cases with sepsis.

Objective To describe the clinical features of invasive fungal disease in Huashan Hospital,Fudan University from January 2004 to December 2006.Methods The medical data were reviewed retrospectively for the patients with fungal infection, which was confirmed by positive fungal culture or microscopic examination with blood,sterile body fluid,deep tissue,sputum specimen or isolation of Aspergillus spp.and Cryptococcus spp.from bronchoalveolar lavage.The proven and probable cases of invasive fungal disease were included in this analysis.Results A total of 111 patients were diagnosed as invasive fungal dis-ease,including 104 proven cases and 7 probable cases.Sixty-one cases were community-acquired and the other 50 were nosoco-mial.The most common site of infection was bloodstream (51,45.9%),followed by central nervous system (44,39.6%)and respiratory system (14,12.6%).The most common pathogens were Candida spp.(50,45%),Cryptococcus (47,42.3%) and Aspergillus spp. (12, 10.8%). The community-acquired fungal infections were mostly found in central nervous system (44,72.1%),and respiratory system (12, 19.7%),mainly caused by Cryptococcus and Aspergillus. The nosocomial fungal infections occurred primarily in blood-stream (96.0%),mainly due to Candida spp.No underlying disease or risk factor was identified in more than half of the pa-tients with community-acquired infection,while almost all the patients with nosocomial fungal infection had underlying disease and predisposing factors.Indwelling venous catheter was closely associated with nosocomial bloodstream infection.Indwelling venous catheter lasted for more than 1 week in 64.7% of the patients with Candida bloodstream infection.The same fungal strain was isolated from both the cather and blood of the same patient in 11 cases.The overall mortality of these invasive fungal diseases was 14.4% (16/111).The mortality rate was 18.0% (9/50)in the patients with nosocomial invasive fungal infection, and 11.5% (7/61)in the patients with community-acquired invasive fungal infection.Conclusions The most common site of in-vasive fungal infection is bloodstream,followed by central nervous system,and respiratory system.Majority of the fungal patho-gens are Candida spp.,Cryptococcus and Aspergillus spp.The community-acquired invasive fungal disease is primarily meningitis caused by Cryptococcus.The nosocomial invasive fungal disease is mainly bloodstream infection caused by Candida spp.

Objective To investigate the correlation of T-lymphocyte expressing HLA-DR with serum HBV DNA and HBeAg contents in chronic hepatitis B. Methods Totally 134 chronic hepatitis B patients and 36 healthy blood donors were enrolled in the study. The T-lymphocytes (CD3 + HLA-DR + ,CD4 + HLA-DR+ and CD8 + HLA-DR+ T) expressing HLA-DR were detected by flow cytometry, the serum HBV viral loads were detected by the real-time quantitative PCR and HBeAg was detected by chemiluminescence method. According to serum HBV DNA viral loads patients were defined as HBV DNA negative (≤ 103 copies/mL), low (＞ 103 - 105 copies/mL), medium (＞ 105 - 107 copies/mL) and high groups (＞ 107 - 109 copies/mL) ; according to serum HBeAg levels, patients were defined as HBeAg negative (≤1 PEIU/mL), low (＞ 1 - 100 PEIU/mL), medium (＞ 100-1 000 PEIU/mL) and high groups (＞ 1 000-10 000 PEIU/mL). T test and one-way ANOVA were performed. Results With HBV DNA loads, HBeAg levels increased, the percentage of CD3 + HLA-DR + , CD4 + HLA-DR + and CD8 + HLA-DR +decreased, especially CD8 + HLA-DR +. Compared with HBV DNA negative group, the percentages of CD3 +HLA-DR + , CD4 + HLA-DR + and CD8 + HLA-DR + were significantly reduced in high group (t = 3. 686,4. 592 and 3. 216, P ＜ 0. 0l); the percentages of CD4 + HLA-DR + and CD8 + HLA-DR + were also reduced in medium group (t = 3. 761 and 2.862, P ＜ 0.01); while in low group, only the percentage of CD8 + HLA-DR + was reduced (t = 2.215, P ＜ 0.05). Compared with HBeAg negative group, the percentages of CD3 +HLA-DR+, CD4 + HLA-DR+ and CD8 + HLA-DR+ were significantly reduced in medium and high groups (thigher =3. 144, 2.222 and 4.035; tmiddle =3.311, 2.362 and 3.374, P ＜0.05), while in the low group,only the percentage of CD8+HLA-DR+ was reduced (t=2.029, P＜0. 05). Conclusion The combined measurement of HBV DNA, HBeAg and T-lymphocytes expressing HLA-DR in chronic hepatitis B patients may not only help to evaluate the immune status of patients, but also can predict the disease progression and clinical outcomes.

0.05),however,it can shorten curative time obviously on Ⅱ type dental ulcer.Simultaneously,the general reaction and focal mucosa of borneolum and borax pellicle group had no obvious variation before and after the test.Conclusions Chitosan has no effect on min-pigs' nerve,cardiovascular and respiratory system;and it is relatively safe given by mouth or peritoneal injection.The borneolum and borax pellicle can shorten curative time obviously on Ⅱtype dental ulcer.

Objective:To explore the strategies and complications of the submucosal tunneling endoscopic resection (STER) in the treatment of esophageal duplicated cyst (EDC).Methods:From January 2013 to December 2019, at Department of Gastroenterology, Taizhou Hospital of Zhejiang Province, the clinical data of 11 hospitalized patients with EDC diagnosed by pathological examination who underwent STER were collected. The clinical featured, computed tomography (CT) findings, endoscopic findings, postoperative efficacy, complications and pathological results after operation were summarized.Results:Among the 11 patients, there were 6 males and 5 females, the age ranged from 13 to 67 years, and the mean age was (43.0±18.2) years. One case presented with swallowing obstruction, 1 case with belching, 4 cases with upper abdominal pain, and the remaining 5 cases had no specific clinical symptoms. Under endoscopy, the lesions of 11 patients were hemispherical or mound-shaped with smooth surface submucosal masses in the esophageal cavity, which were soft to touch. Under endoscopic ultrasonography, they all appeared as hypoechoic masses with clear boundary located in the esophageal muscularis propria. The results of CT scan of 9 patients showed round low-density cystic masses, among them 7 cases showed mild enhancement. The maximum diameter of the cysts was 1.5 to 4.4 cm, with mean maximum diameter being (2.8±0.8) cm, and the maximum diameter of most patients (7 cases) were 2 to 3 cm. The other two patients showed only slightly thickened esophageal wall on CT. Five lesions occurred in the horizontal mediastinum of the lower esophagus. All the 11 patients underwent STER operation successfully, among them 6 patients received simple STER and the cysts were completely resected, and the other 5 patients received STER and cauterization with argon ion for the residual cyst wall. The pathological results after operation indicated that 6 cases were congenital esophageal cysts and 5 cases were bronchogenic cysts. The median follow-up time (range) of 11 patients was 42 months (12-86 months). Ten patients recovered well after the operation, and local recurrence, malignant transformation or metastasis were not found. One case had recurrence, and was treated with STER and cauterization with argon ion for residual cyst wall and cured. No complications such as bleeding, fistula, mediastinal infection or death occurred during and after operation in all the 11 patients.Conclusions:STER is a minimally invasive, safe and effective treatment for EDC, and may be a new treatment for EDC.

Objective: To dynamically evaluate left ventricular perfusion, global and local functional changes during left ventricular aneurysm (LVA) formation and to explore the relationship between the size of LVA and LVEF, LVESV, LVEDV by gated99mTc-MIBI SPECT (GSPECT) and gated18F-FDG PET metabolic (GPET) imaging in experimental pigs. Methods: LVA model was established by occlusion of left circumlfex artery (LCX) and placing an Ameroid constrictor at the proximal end of left anterior descending artery (LAD) in a total of 16 Chinese mini-pigs. At the 1st, 4th and 8th weeks of surgery, the changes of total perfusion defect (TPD), LVA formation and LVEF, LVESV, LVEDV were dynamically evaluated by GSPECT and GPET; the relationships between the size of LVA and LVEF, LVESV, LVEDV were analyzed respectively.Results: There were 5 pigs died in surgery and 2 died at the 1st week of modeling. According to golden (pathological) standard, 9 animals successfully ifnished the dynamic imaging study. At the 1st week of (basic) modeling, 4 animals formed large LVA, 2 formed small LVA at the apex and 3 without LVA formation. At the 4th and 8th weeks of modeling, dynamic imaging presented that the animals with large LVA had gradually increased range and degree of perfusion defect, LVEDV, LVESV, while gradually decreased LVEF; the above indexes were relatively stable in animals with small or none LVA. In addition, the size of LVA was related to LVEF (r=-7.26), LVEDV (r=0.855) and LVESV (r=0.825), allP<0.05. Conclusion: In experimental pigs, at the beginning of LVA formation, large range and severe perfusion defect may cause large aneurysm, the LV functional damage and remodeling may gradually increase and the prognosis is poor; in contrast, the animals with small or none LVA have better prognosis and usually without ventricular remodeling; which implies that in acute phase of LVA formation, the size of aneurysm may predict the trend of global LV systolic function and remodeling at the early stage.

Objective To understand the genotypic characteristics and the neutralizing antibody levels of Japanese encephalitis virus(JEV)and Japanese encephalitis(JE)in both vector mosquitoes and in healthy people of Zhejiang province. Methods Virus was isolated from mosquitos sampled from the Monitoring Stations located in Xianju county during 2012 to 2013. Phylogenetic and homological studies were carried out on the E gene. A total of 1 263 blood specimens from 642 healthy people were collected before and after the seasons of JE epidemics. JEV neutralizing antibody was detected by the micro-neutralization test. Results Twenty-five JEV strains were isolated from a total of 11 650 mosquitoes. The identity of nucleotide appeared as 87.8%-99.7% both from 2012 to 2013 and from 1982 to 2010 while as 87.7%-88.0%with vaccine strain SA14-14-2,in Zhejiang. The phylogeny tree of E gene indicated that the newly isolated virus belonged to genotypeⅠ but no mutation of amino acid sequence coding conformational epitope was identified in the envelop protein. Both positive rates and the geometric mean titer(GMT)of neutralizing antibody in healthy people were 31.5%-42.0%and 1∶2.56-1∶3.53 in Xianju county,during 2012 and 2013,respectively. Both of the two positive rates(χ2≤1.76,P>0.05)and the two GMTs(u≤0.64,P>0.5)for antibodies pre or post the epidemic season did not show significant differences. Conclusion JEV isolated in Xianju during 2012 and 2013 belonged to genotypeⅠ. The positive rates of JEV neutralizing antibody from healthy people in Xianju were less than 42.0%,which showed no significant differendes pre or post JE epidemic season.

Objective To investigate the level of CD31 molecule expression on peripheral T lymphocytes of patients with chronic hepatitis B (CHB),and to study the relationship between the expression of CD31 molecule and serum ALT levels,serum levels of HBV DNA and HBeAg.Preliminarily explore the relationship between the expression frequency of CD31 molecule and patients CHB,in order to understand changes in different immune status when chronic HBV infecting.Methods A total of non-antiretroviral therapy 42 patients with CHB and 25 healthy volunteers were enrolled for the study.In peripheral blood,separated lymphocytes conventionally,the percentage of CD4 + 、CD8 + 、CD31 + 、CD31 + CD4 + 、CD31 + CD8 + 、CD31 + CD4 +/CD4 + 、CD31 + CD8 +/CD8 + cells were assessed by flow cytometry,serum ALT levels were tested by automatic biochemical analyzer,serum HBVDNA levels were determined by real-time fluorescent quantitative PCR,and the serological markers of HBV were examined through chemiluminescence simultaneously.Results The percentages of CD4 + 、CD31 + 、CD31 + CD4 + 、CD31 +CD4 +/CD4 + cells in patients with CHB were significantly less than the healthy(P ＜ 0.05);the percentages of CD31 + 、CD31 + CD4+ 、CD31 + CD8+ 、CD31 + CD4+/CD4+ cells in ALT ＜2ULN group of patients with CHB were significantly lower than ALT≥2ULN group(P ＜ 0.05);serum HBVDNA loads lower group (＞103-105copies/ml),middle group (＞ 105-107 copies/ml) and higher group of HBVDNA (＞ 107-109 copies/ml) CD31 molecule expression frequency were on the rise,but no statistically significant difference were observed in the comparison between groups (P ＞ 0.05);the percentages of CD31 + 、CD31 + CD8 + 、CD31 + CD4 +/CD4 + cells in the HBeAg-negative group were significantly lower than HBeAg-positive group (P ＜0.05).Conclusions The frequency of CD31 molecule expression on peripheral T lymphocytes of patients with CHB were declined;there is a certain relationship between the frequency of CD31 molecule expression and serum ALT levels,different loads of HBVDNA,HBeAg positive or negative in patients with CHB;chronic HBV infection of immune status may be associated with T cell surface expression of CD31 molecule frequency.

Objective: To assess the feasibility of coronary lfow reserve (CFR) detection by SPECT myocardial perfusion imaging using a self developed software with preliminary clinical veriifcation. Methods: CFR calculation software was developed according to Mat lab guide. A total of 16 patients were enrolled including 13 male and 3 female at the mean age of (58±11) years . CAG conifrmed that 25 coronary branches were with stenosis>50% and 23 branches were without stenosis. 2-day ATP/rest99mTc-sestamibi dynamic SPECT myocardial perfusion imaging was conducted to detect CFR. First transit counts were used to sketch the interested pulmonary artery segments and to obtain the arterial input curve of contrast agent as total PAC reached to heart. Reconstructed short-axis images were divided into 3 sections to sketch interested territories (ROI) and to obtain RMC at each territory. Estimated CFR was expressed by the ratio of MBF=RMC/PAC followed by calculating the ratio of MFR=MBFstress/MBFrest. Results: The difference between simulated value and true value could be ignored which conifrmed that our program may accurately measure CFR. The reproducibility by different operators (r=0.986) and the same operator (r=0.983) was good. CFR value in non-stenosis branches were higher than stenosis branches (1.28 ± 0.19) vs (1.10 ± 0.27),P=0.008 and CFR value in stenosis branches was negatively related to stenosis degree (r=-0.5,P=0.02). Conclusion: Our self developed software is reliable for CFR detection by SPECT myocardial perfusion imaging; preliminary study showed good application prospect in clinical practice.