The synchronous fluorescence spectra of myoglobin were studies for the first time. The fluorescence peaks observed in the spectra were assigned. When the wavelength interval (Δλ) is 80 nm, the main peak at 335 nm is originated from the tryptophan residues in the myoglobin molecule. When Δλis 20 mn, the peak at 308 nm is mainly due to the tyrosine residues in the myoglobin molecule and in a small part due to the tryptophan residues.Two peaks at 322 and 596 nm were observed in the spectrum of myoglobin for Δλ = 40 nm. The peak at 322 nm is due to both tyrosine and tryptophan residues. The peak at 596 nm is attributed to the heme group in the myoglobin molecule .

Soft-tissue deformation is one of the important research directions in virtual surgery. Mass-spring model and finite-element model are the two most important modeling technology of the soft tissue modeling. It is very important for the current soft-tissue deformation modelling to analysize, to summarize the main idea, the influence factors and the modeling route of the two kinds of modeling methods and then to give comprehensive review.
Computer Simulation ; Humans ; Models, Biological ; Surgery, Computer-Assisted ; methods ; User-Computer Interface

Objective To determine the differentially expressed genes in the development of vascular medium calcification in rats using the suppression subtractive hybridization (SSH). Methods Twenty-four 6-week old SD rats of specific pathogen free grade were recruited and randomly allocated into calcified group (n=12) and control group (n=12). Rats were made for vascular calcification model in calcified group (vitamin D3 plus nicotine, VDN). All rats were sacrificed to measure concentration of calcium in the arterial tissue and examine the pathological lesion changes. RNA in rat aortic tunica tissue was extracted and reverse transcripted into cDNA. cDNA fragments which highly expressed calcification were isolated in calcified group using the SSH. Differentially expressed genes with cDNA fragment were inserted into PMD18-T plasmid vector and transformed to competent DH-5α by means of heating transfer. cDNA libraries of differentially expressed gene between calcified group and control group were successfully constructed. Recombinant vectors were analyzed by colony PCR. Positive genes were randomly selected for sequencing and analyzed by BLAST. Six genes, for example, were randomly selected for RT-PCR certification. Results (1) The pathological examination results demonstrated that in calcified group there were obvious calcium diposits and media squirm in tunica media of rat aortic wall, while in control group no calcium diposit was found. (2) There was statistical significance in calcium concentration in vascular tissue between calcified group[(15.34 ± 2.51)mg/g] and control group [(5.20 ± 0.75) mg/g] (P＜0.01). (3) Subtracted libraries in vascular calcification was successfully established. Ninety-two positive clones in positive library and 18 positive clones in reverse library were obtained after the colony PCR identification. The length of insertion fragments was concentrated between 150 bp and 400 bp. Calcification-related 43 up-regulated genes and 11 down-regulated genes were obtained through sequencing and BLAST analysis in positive clones. RT-PCR validation indicated that the expressions of 5 genes such as CytoP450 and Nell1 had greater increase in calcified group than those in control group, the average fold change was 1.71.Conclusions Model of vascular calcification induced by vitamin D3 plus nicotine is successfully constructed. Related gene expression spectrum is changed in the process of vascular calcification.Some ossification genes and genes associated with apoptosis, oxidation, inflammation and cytokines are up-regulated. At the same time, some genes which possibly inhibit vascular calcification are down -regulated.

Objective To observe any therapeutic effect of laser irradiation on pathological inflammatory reactions in ulcerative colitis (UC) induced by oxazolone,and to investigate possible mechanisms.Methods Six rats were selected as a normal control group.Another 24 rats with UC induced by oxazolone were randomly assigned to a UC model group (n =8),a 400 mW laser group (161.3 mW/cm2,n =8),and a 200 mW laser group (80.6 mW/cm2,n =8).All the rats were fixed in custom-built devices.Those in the therapy groups were treated daily,10 min per time,for 10 days.After the end of the last irradiation session,disease activity indexes (DAIs)were observed.Rats from every group were sacrificed 24 h after the last irradiation in order to observe any pathological changes in colon tissue,the weight of fresh ulcerated tissues,and gross changes in morphology.Colon segments were stained with hematoxylin-eosin and histological lesion scoring was performed under a light microscope.Any changes in inflammatory edema in the colonic mucous membrane were observed before and after laser irradiation.Results The UC model was successfully established.Average body weight in the 400 mW laser group increased significantly more than in the UC model group,approaching that of the normal control group.DAI decreased significantly.The thickness of the epithelial mucous membrane,lamina propria and submucosa was basically restored.Histological lesion scores also improved significantly,and the weight of fresh ulcerative tissue was significantly lower.Mucous membrane ulcers,submucosa edema and inflammatory cell infiltration all were alleviated significantly,merely presenting a few inflamed cells and small amounts of periphlebitis.In the 200 mW laser group all these outcome measures improved significantly compared with the UC model group,but were not as good as in the 400 mW laser group,and the rats needed longer to recover.Conclusion Laser irradiation at 400 mW has advantages over 200 mW,and could significantly relieve the pathological inflammatory response in colonic tissue,decrease submucosa edema and ameliorate other symptoms of ulcerative colitis,at least in rats.

Objective To explore the effects and molecular mechanisms of laser therapy on serum and colon tumor necrosis factor α (TNF-α),interleukin-6 (IL-6) and interleukin-10 (IL-10) in rats with oxazolone induced ulcerative colitis (UC).Methods Thirty adult male SD rats were randomly divided into four groups:a normal group ( n =6),a UC model group ( n =8 ),a 400 mW laser treatment group ( n =8 ) and a 200 mW laser treatment group ( n =8 ).Odified oxasolone sensitization was used to induce UC models in the rats.The AsAlGa semiconductor laser used in the treatment had a power of 200 mW or 400 mW.The therapy lasted for 10 days with daily 10 min sessions.The rats were sacrificed after treatment and enzyme-linked immunosorbent assay (ELISA) was used to measure IL- 6,IL- 10 and TNF-α in serum and ulcer tissues in all groups.Results Compared with the normal group,the weights of the UC model rats were significantly lower,and they had severe mucoid bloody stool.TNF-α and IL-6 levels in serum and colon tissues increased significantly and IL-10 decreased significantly,so the UC model was successfully established.After laser treatment,body weight gain and stool moderation in rats were observed.in the 400 mW group,and average TNF-α and IL-6 levels in both serum and colon tissue decreased significantly.IL-10 increased significantly,close to the level of the normal group.In the 200 mW group,serum TNF-α and IL-6 levels were significantly lower,and IL-6 in the colon tissue was reduced significantly.TNF-α did not lessen significantly,and serum and colon tissue IL-10 levels did not improve significantly compared with the model group.Conclusions 400 mW AsAlGa semiconductor laser irradiation can effectively regulate cytokines after oxaZolone induced UC in rats in two ways.It reduces pro-inflammatory cytokines and increases the level of anti-inflammatory cytokines.This may imply one of the underlying molecular mechanisms of low energy laser treatment of UC.

A total of 169 patients with short-duration type 2 diabetic mellitus (DM) were divided into atherosclerosis (AS) group and non-AS group according to the intima-media thickness (IMT) of three conducting arteries.The level of serum amyloid A (SAA) was assayed by ELISA.The results showed that SAA level of type 2 DM patients increased significantly, patients in AS group showed higher SAA level than that in non-AS group, and SAA level was positively correlated with age, body mass index, waist hip ratio and IMT of common carotid artery.Age, C-reactive protein and SAA level are the major risk factors for IMT of common carotid artery.

Objective:To assess the validity and reliability of the Chinese version of the eleven-item Kutcher Adolescent Depression Scale (KADS-11)in Chinese adolescents,calculate its optimal cut-off value and the sensi-tivity and specificity,and explore the possibility of providing a useful tool to assess the severity of adolescent de-pressive symptoms.Methods:Totally 3180 students aged 11 -17 years were selected from schools in 6 provinces and Shanghai.All of them were asked to complete the KADS-11 and Children Depression Inventory (CDI). Students whose CDI scores were above 19 (including 19)were diagnosed with the DSM-IV criteria of depressive disorder,73 students from Shanghai sample were assessed with KADS-11 and CDI to analyze the test-retest reliabil-ity 1 month later.Results:Exploratory factor analysis showed that KADS-11 had 2 factors,and confirmatory factor analysis tested proved the 2-factor model fit better than the one-factor model.The KADS-11 total scores were posi-tively correlated with CDI total scores (r =0.74,P <0.01 ),and the KADS-11 scores were higher in depressive group than those in non-depressive group.The mean area under the curve (AUC)of KADS-11was 0.94,the mean area under the curve of each item ranged from 0.7 to 0.9.The optimal cut-off point of KADS-11 was total score≥9,sensitivity and specificity were 89% and 90% respectively.The Cronbach's alpha coefficient of the KADS-11 was 0.84,the spilt-half reliability coefficient was 0.71 (P <0.01),and the test-retest coefficient was 0.77 (P <0.01).Conclusion:The KADS-11 is appropriate for Chinese adolescents because of its good validity,reliability and diagnosis accuracy,it could be used to assess depressive symptoms for adolescents.

OBJECTIVETo investigate the in vitro effects of immune inhibitor tacrolimus on platelet function.

METHODSFresh venous blood was collected from healthy volunteers at ages of 18-25 years old, who are not taking antiplatelet drugs within two weeks. The platelets were isolated from the blood and incubated with different concentrations of tacrolimus (0.06, 0.6, 6, 60, 120, 240 μmol/L) at 37 °C for 2 hours, and then the changes of mitochondrial membrane potential and P-selection of platelets were detected by flow cytometry, the expression of apoptosis related protein by Western Blot, and the change of the platelet aggregation function by platelet aggregation analyzer.

RESULTSTacrolimus at concentration of 0.06 μmol/L could promote collagen induced platelet aggregation, inhibit thrombin induced platelet aggregation, have no effect on ristocetin and vWF induced platelet aggregation function. Tacrolimus at concentration of 120 μmol/L and 240 μmol/L could reduce the platelet mitochondrial membrane potential and induce the expression of apoptosis protein caspase-3.

CONCLUSIONIn vitro experimental results showed that high concentration of tacrolimus could lead to platelet apoptosis. But the current therapeutic dose of tacrolimus at 0.06 μmol/L (which is equivalent to 50 ng/ml blood concentration) could have different effects on platelet aggregation function according to different stimulating agents.

Adolescent ; Adult ; Blood Coagulation Tests ; Blood Platelets ; drug effects ; Caspase 3 ; Humans ; In Vitro Techniques ; Platelet Aggregation ; Tacrolimus ; pharmacology ; Thrombin ; Young Adult

OBJECTIVETo establish a rapid, accurate, noninvasive and low cost method for screening MT3243A>G mutation in mitochondrial diabetes.

METHODSBlood, saliva, and urine sediment samples were collected from 6 patients with confirmed mitochondrial diabetes and 50 healthy controls from Shanghai Children's Hospital and Shanghai Sixth People's Hospital. The heterozygosity levels of MT3243A>G mutation in above samples were detected with pyrosequencing, and the data were compared. MT3243A>G mutations were rapidly screened with high resolution melting curve analysis (HRM) in the urine sediment samples of 1070 diabetic patients from 4 communities in Shanghai. Furthermore, pyrosequencing was used to validate the suspected positive samples, and the heterozygosity levels were also quantified.

RESULTSComparative experiments found that heterozygosity of MT3243A>G mutation was 2 to 7 times higher in urine sediment than in saliva and blood samples from the 6 patients with confirmed mitochondrial diabetes. However, the heterozygosity was slightly higher in saliva than blood samples. MT3243A>G mutation was not detected in the 50 healthy controls. Two samples with suspected MT3243A>G mutation were identified in the 1070 urine sediment samples of diabetes patients with HRM screening, which were validated by pyrosequencing. The heterozygosity of MT3243A>G mutation were 33.32% and 14.67% in the urine sediment samples, respectively.

CONCLUSIONUrine sediment samples can be used for rapid screening of MT3243A>G mutation for its ease to collect, noninvasiveness and higher level of heterozygosity. HRM is suitable for rapid screening for mitochondrian mutations for its low cost, while such mutations could be detected with sensitivity and accuracy by pyrosequencing.

DNA, Mitochondrial ; genetics ; Diabetes Mellitus ; genetics ; Heterozygote ; Humans ; Mutation ; Sequence Analysis, DNA ; methods ; Transition Temperature

【Objective】 To track and evaluate the clinical diagnostic efficacy of an enzyme linked immunosorbent assay (ELISA) kit for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). 【Methods】 Total antibody (TAb) specific to SARS-CoV-2 in blood donors were determined using ELISA reagent. TAb positive donors were followed up 1 month after blood donation. SARS-CoV-2 specific IgG, IgM and pseudotype lentivirus based neutralization test (ppNAT) were conducted for TAb positive blood donors and follow-up samples. ppNAT and IgG antibodies simultaneously positive in ppNAT positive samples and its follow-up samples was used as the standard for antibodies validation. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), accuracy and Youden index of SARS-CoV-2 TAb ELISA were analyzed. 【Results】 Among 16 016 blood donors from January 31 to April 28, 2020, 61 donors were diagnosed as TAb positive, 6 cases were positive for ppNAT, in which 2 were positive for both ppNAT and IgG; 4 of 46 TAb positive follow-up samples were positive for ppNAT, in which 2 were positive for IgG simultaneously. The sensitivity, specificity, PPV, NPV, accuracy, Youden index, false positive rate and false negative rate of SARS-CoV-2 TAb reagent were 100.00%, 99.60%, 3.28%, 100.00%, 99.60%, 99.60%, 0.40% and 0.00%, respectively. 【Conclusion】 SARS-CoV-2 TAb ELISA has high sensitivity and good clinical diagnostic efficacy, but the false positive rate is relatively high in low-risk blood donors. Therefore, ppNAT, IgG and follow-up results should be fully considered in clinical in order to analyze the positive results and determine the infection status more accurately.