ObjectiveTo explore the relationship between the T3394C mutation of the mitoehondrial DNA ND1 gene and type 2 diabetes mellitus (T2DM) in the elderly. MethodsDirect sequencing of PCR product was applied to study the T3394C mutation of mitochondrial DNA in 340 unrelated individuals in T2DM patients (90 cases of elderly patients, 250 cases of non-elderly patients) and 265 normal controls (130 cases of elderly controls, 135 cases of non-elderly controls) from Zhejiang province. The mutation was analyzed by DNASTAR and Antheprot 5.0 softwares. ResultsThe T3394C mutation of mitochondrial DNA was found in 5 of 90 elderly T2DM patients (5.6%),1 of 130 elderly controls(0.8%), and 2 of 250 non-elderly T2DM patients (0.8%). The incidence of the mutation was significantly different between the elderly T2DM patients and elderly controls(P＜0.05), as well as between the elderly T2DM patients and non-elderly T2DM patients(P＜0.05). The T3394C mutation caused the change of the ND1 protein secondary structure. ConclusionsThe T3394C mutation of the mitochondrial DNA ND1 gene may contribute to the pathogenesis of T2DM in the elderly with other genetic and environment factors.

AIM: To study the immunity modulation mechanism of achyranthes bidentata polysaccharides (ABPS). METHODS: (1) PBMC from patient with asthma or lung cancer were cultured with 800 mg/L ABPS for 18 h. IFN-? and IL-4 mRNA was detected by RT -PCR. (2) Th cells were cultured with serially diluted ABPS for 48 h and with 800 mg/L ABPS for various times. Cultured supernatant was harvested at diffevent time points and RNA was extracted, respectively. IFN-? or IL-4 transcriptional and translational levels were assay by RT -PCR and ELISA. RESULTS: (1) Positive rate of IFN-? mRNA expression in PBMC from patient with asthma or lung cancer increased from 6/25 to 14/25 (P

To elucidate the signal transduction mechanism of systemic inflammatory response syndrome induced by LPS,Western blot analysis was used to assay the protein expression and phosphorylation levels of ERK, JNK,and p38 MAPK in murine KC which were stimulated by LPS. It was found that LPS treatment resulted in no significant change in their protein expression, but a rapid transient increase in their phosphorylation levels, which peaked at 30, 45,and 20 minutes,respectively; The values returned to mear baseline within 2 hours. Kinase phosphorylation levels of these three MAPKs induced by LPS were found to be dose dependent in a range of 10 pg/ml to 100ng/ml. No phosphorylation was observed in unstimulated cells. Among them, p38 MAPK shoned fastest and highest phosphorylation levels. The results show that LPS can markedly activate ERK, JNK,and p38 MAPK in KC, and p38 MAPK may play a more important role in LPS induced kupffer cell activation than ERR and JNK.

Objective To explore the effects of interleukin- 1?(IL-1?)on the biological activity of human periodontal ligament(PDL) cells in vitro. Methods Human PDL cells were cultured in DMEM medium containing IL-1?(0.1,0.5,1,5 and 10ng/ml) for 1,2,3,4 and 5 days, respectively. The proliferation of PDL cells was measured by MTT assay at the first, second, third, fourth and fifth days after IL-1? treatment, respectively. Fibronectin level in the medium was determined by ELISA at the fourth days after 10ng/ml IL-1? treatment, and alkaline phosphatase(ALP) activity was measured by enzyme kinetic method. Results IL-1? inhibited the growth of human PDL cells in a dose-dependent manner, and its lowest effective concentration was 1ng/ml(P

Objective To investigate clinical value of the level of arterial plasma colloid osmotic pressure (COP) in predicting the prognosis of severe patients.Methods According to the prognosis of patients,the patients were divided into survival group and death group,12 h arterial plasma COP change level in the two groups were compared.According to APACHE Ⅱ score,the patients were divided into≤25 and ＞25 group,the COP into the ICU,12 h plasma COP change were compared.The correlation of APACHE Ⅱ score and arterial lactate into ICU,12 h arterial plasma COP were analyzed by Spearman correlation analysis.Results The level of plasma COP into the ICU and 12 h levels of survival were significantly lower than those of the death group; in the APACHE Ⅱ score≤25 group,plasma COP into the ICU levels and 12 h plasma COP levels were significantly higher than those of the APACHE Ⅱ score ＞ 25 group,and 12 h COP levels were significantly higher than those of behind group (t =8.34,P ＜ 0.01),Further analysis showed that the COP level into the ICU and APACHE Ⅱ score was significantly negative correlated; 12 h COP levels and APACHE Ⅱ score was significantly negative correlated.Conclusions Dynamic monitoring of COP levels is a good indicator to determine the prognosis of patients with the danger patients,the lower plasma COP,the more severe disease and poor prognosis.

Objective To investigate the effect of deferasirox on DLL4 expression and angiogenesis in a narrow pedicle and random flap in rats.Methods Twenty SD rats were randomly divided into the deferagirox group and control group.Rats were subjected to deferagirox of 100 mg · kg-1 · d-1 inthe experimental groups,respectively and the same dose saline in the control group for 1 week. In each group,flap were created in the bilateral back of each rats.Ratio of length to width of tissue in the pedicle portion and the flap portion was 1 cm × 1 cm and 3 cm × 3 cm,respectively.The tissue samples were taken from the pedicle and the middle portions of the flap.The DLL4 and CD105 expression was also detected with immunohistochemistry (SABC).Results Compared with control group,whatever in the pedicle portion or the middle portion,there was a significant increase of microvessels marked by CD105 and a significant decrease of flap microvessels stained by DLL4 in the deferagirox group.In both groups,compared with the pedicle portion,there was a significant increase of microvessels marked by CD105 and DLL4 in the the middle portion.Conclusions Deferasirox can in crease the CD105 expression and angiogenesis of the slender narrow pedicle random flap.This process might be related to the inhibition of DLL4 protein expression,which is significant in the notch signaling pathway.

Objective To analyze the specific peptide of cystatin C (CysC) and its characteristics by bioinformatics technology,and verify the predicted results by mass spectrometry.Methods Online software was applied to analyze the physicochemical properties and homology of CysC peptides hydrolyzed by trypsin and predict the associated parameters of ionized fragmentation of specific peptide by mass spectrometry.Precursor ion scan and product ion scan were conducted on the samples of synthetic specific peptide.The recombinant human CysC and serum samples were analyzed by mass spectrometry after trypsin digestion.The results of analysis were compared with the outcomes predicted by bioinformatics.Results T3 (ALDFAVGEYNK) was considered as the specific peptide of CysC by software analysis.When selecting[M + 2H] 2 + for product ion scan,almost all the y and b ions of fragmentation were observed using tandem mass spectrometry (MS/MS),showing consistency with Skyline predictions.Moreover,both the peptides from the human recombinant CysC and serum sample following the trypsin digestion were eluted at the same time with the isotope-labeled T3 * under the fixed conditions.Conclusion Bioinformatics technology could be available for picking out the specific peptides of target protein quickly and efficiently and predicting the ionized fragmentation precisely by mass spectrometry scanning.

Objective To establish the methodology of uncertainty evaluation with reference measurement procedure by Monte Carlo method (MCM).Methods According to JCGM 101:2008,we established the methodology of uncertainty evaluation by MCM in the example of GGT reference measurement procedure.We could calculate an estimate of GGT concentration,the associated standard uncertainty and a coverage interval with a specified probability by MATLAB software,setting the uncertainty evaluation by GUM method as a control.Results When the uncertainty was evaluated by GUM method,the results of sample A and Sample B were (95.8 ±2.4) U/L (k =2) with coverage interval [93.4 U/L,98.2 U/L] and (180.0 ± 3.9) U/L (k =2) with coverage interval [176.1 U/L,183.9 U/L] respectively,while using MCM method,the uncertainty evaluation result of sample A and Sample B were (95.8 ± 2.4) U/L (k =2) with coverage interval [93.4 U/L,98.2 U/L] and (180.0 ± 3.9) U/L with symmetrical 95％ coverage interval [176.2 U/L,183.8 U/L].The output quantity simulated by MCM was normal distributed.Conclusions When the distribution of the output quantity is normal,the measurement uncertainty evaluated by both MCM and GUM method is nearly the same.When the distribution of the output quantity is unknown,MCM can be used as a verification of GUM method.

Background and purpose:Multiple primary lung cancers (MPLC) is a rare entity, but recently there has been a gradual increase in the number of patients diagnosed with MPLC. The aim of this study is to investigate the diagnosis, treatment and prognosis of MPLC through analyzing the clinical data.Methods:Forty-one patients were diagnosed MPLC by Martini-Melamed criteria. Their clinicopathological data were retrospectively reviewed. Results:There were 3 patients with triple primary lung cancer and 38 patients with double primary lung cancer. There were 13 patients with synchronous MPLC, 26 patients with metachronous MPLC, 2 patients with synchronous and metachronous MPLC. Of 85 lesions, the surgical procedures were mainly lobectomy (78.8%, 67/85). Lesions (41.2%, 35/85) were frequently in right upper lobe. Pathological type was mainly adenocarcinoma (70.6%, 60/85),followed by squamous cell carcinoma (17.6%, 15/85). Of 60 adenocarcinoma specimens, the papillary predominant subtype was more common (50%, 30/60). Eighty percent (68/85) of the lesions were stage I. As to the initial cancer and repeated cancer, patients who shared the same pathological type (68.3%, 28/41) were more than the different (31.7%, 13/41), of which adenocarcinoma-adenocarcinoma was most common(82.1%, 23/28). Lesions located in contralateral lobes were in 37 patients (90.2%), and located in ipsilateral different lobes were in 4 patients (9.8%). The 2-year overall survival (OS) of them was 87.8%. Survival analysis showed that the prognosis of patients with same pathological type was better than patients with different pathological type (P=0.037), the prognosis of patients with no lymph node metastasis was better than patients with N1,N2 metastasis (P=0.02).Conclusion:Lesions in patients with multiple primary lung cancers are more frequently in the right upper lobes. The pathology type is mainly adenocarcinoma, of which the papillary predominant subtype was most common. Early diagnosis improves continuously, active treatment with operation can achieve better prognosis.

Objective:To evaluate the clinical feasibility and image quality of three-dimensional breath-hold gradient and spin-echo (3D BH-GRASE) sequence in magnetic resonance cholangiopancreatography (MRCP).Methods:In this prospective study, 59 patients with clinically suspected pancreaticobiliary duct disease performed MRCP with both 3D BH-GRASE and 3D respiration-triggered turbo spin-echo (3D RT-TSE) sequences on 3.0 T scanner in the Affiliated Zhangjiagang Hospital of Soochow University from November 2017 to December 2018. The overall image quality was scored independently by 3 experienced radiologists based on the visibility of different anatomical features of the pancreaticobiliary duct. For comparing the 2 sequences, the statistical difference in scan time was assessed with a paired t test; while subjective scores, signal-to-noise ratios (SNR), contrast ratios (CR) and contrast noise ratios (CNR) were compared with Wilcoxon signed rank test. Results:The scan time of 3D BH-GRASE sequence was 16.4 s while that of 3D RT-TSE was (258.6±42.2) s. Their difference was statistically significant ( t=44.073, P<0.001), with the scan time for 3D BH-GRASE shortened by 94%. The overall quality scores of 3D BH-GRASE images were better than those of 3D RT-TSE ( Z=-6.595, P<0.001). There was no statistical difference ( P>0.05) in the scores regarding the visibility of the upper, middle and lower parts of common bile duct and the first and second branches of left and right hepatic ducts. For visualizing the bottom, body, neck and duct of gallbladder, the 3D BH-GRASE sequence received a higher score than the 3D RT-TSE sequence ( P<0.001). For displaying the proximal, middle and distal segments of main pancreatic duct, the 3D RT-TSE sequence was scored higher than the 3D BH-GRASE sequence ( P<0.05). There was no significant difference of SNR between the two sequences ( Z=0.403, P=0.687), whereas CR and CNR of 3D RT-TSE MRCP were better than those of 3D BH-GRASE MRCP ( Z=6.215, P<0.001 and Z=3.046, P=0.002, respectively). Conclusion:Under the prerequisite of ensuring image quality, a proper use of 3D BH-GRASE sequence can significantly shorten the scan time and thus greatly improve the working efficiency of MRCP examination.