abstract Cell metabolite analysis is of great interest to analytical chemists and physiologists, with some meta-bolites having been identified as important indicators of major diseases such as cancer. A high-throughput and sensitive method for drug metabolite analysis will largely promote the drug discovery industry. The basic barrier of metabolite analysis comes from the interference of complex components in cell biological system and low abundance of target substances. As a powerful tool in biosample analysis, microfluidic chip enhances the sensitivity and throughput by integrating multiple functional units into one chip. In this review, we discussed three critical steps of establishing functional microfluidic platform for cellular metabolism study. Cell in vitro culture model, on chip sample pretreatment, and microchip combined detectors were described in details and demonstrated by works in five years. And a brief summary was given to discuss the advantages as well as challenges of applying microchip method in cell metabolite and biosample analysis.

Objective:To investigate the correlation of serum testosterone level with severity and characteristics of coronary plaque, stent implantation rate and major cardiovascular adverse events(MACE)in elderly male patients with coronary heart disease(CHD).Methods:In this retrospective study, a total of 63 elderly male patients of the Third People's Hospital of Hangzhou with coronary angiography(CAG)-confirmed CHD and to undergo percutaneous coronary intervention(PCI)were selected.According to serum testosterone level, they were divided into the low testosterone(low T)group and the normal testosterone(normal T)group.Optical coherence tomography(OCT)was performed in both groups to define the characteristics of coronary artery lesions and guide stent implantation.The correlation of serum testosterone level with blood lipids, glycated hemoglobin(HbA1c), degree of coronary artery lesions, plaque characteristics, stent implantation and MACE in two groups were analyzed.The in-stent restenosis rate after stent implantation and the variation of minimum lumen diameter of stent were determined during 12 months follow up in both groups.Results:Total cholesterol(TC), low-density lipoprotein(LDL-C)and HbA1c were higher in the low T group than in the normal T group( t=7.808、-5.871、6.611, all P<0.05). When taking testosterone as the independent variable, and TC, triglycerides(TG), LDL-C, high density lipoprotein cholesterol(HDL-C)and HbA1c as the dependent variables, linear regression analysis showed that TC, LDL-C and HbA1c were negatively correlated with testosterone level( β=-0.733, -0.716, -0.581, P<0.05). More than 2 vascular lesions were more common in low testosterone group versus the control group( χ2=8.66, P<0.05). Mixed plaques, lipid plaques, and calcified plaques were more commonly found in low testosterone group versus the control group( χ2=7.87, P<0.05). Unstable plaques were more common in the low T group( χ2=6.14, P<0.05). The low T group vs the normal T group, coronary stent implantation rate were 93.3%(28/30 cases) vs.66.7%(22/33 cases), the difference was statistically significant( χ2=6.82, P<0.05). When testosterone, TC, TG, LDL-C, HDL-C, HbA1c were taken as the independent variables, and the stent implantation rate was the dependent variable, logistic regression analysis results showed that only testosterone, TC and HbA1c were independently correlated with stent implantation rate( OR=0.971、425.523、0.004, P<0.05). There was no statistically significant difference in minimum stent lumen diameters between the two groups under OCT-guided coronary stent implantation( t=-1.064, P>0.05). During 12 months follow up, the MACE0 incidence was 26.7%(8/30 cases, in low T group)than 6.1%(2/33 cases, in normal T group), with statistically significant difference( χ2=5.00, P<0.05). When taking testosterone, TC, TG, LDL-C, HDL-C and HbA1c as the independent variables, and MACE as the dependent variable, logistic regression analysis results showed that only testosterone and LDL-C were independently correlated with MACE( OR=0.968, 0.008, P<0.05). Conclusions:Serum testosterone level is negatively correlated with TC, LDL-C and HbA1c, and may be correlated with the degree of coronary artery lesions, plaque properties, MACE and stent implantation rate of CHD patients.Serum testosterone can be used to evaluate the characteristics and conditions of CHD, and help to predict the prognosis of CHD.The OCT is a good guide tool for coronary stent implantation.

Livestock husbandry is vital to economy of the Tarim Basin, Xinjiang Autonomous Region, China. However, there have been few surveys of the distribution of ixodid ticks (Acari: Ixodidae) and tick-borne pathogens affecting domestic animals at these locations. In this study, 3,916 adult ixodid ticks infesting domestic animals were collected from 23 sampling sites during 2012-2016. Ticks were identified to species based on morphology, and the identification was confirmed based on mitochondrial 16S and 12S rRNA sequences. Ten tick species belonging to 4 genera were identified, including Rhipicephalus turanicus, Hyalomma anatolicum, Rh. bursa, H. asiaticum asiaticum, and Rh. sanguineus. DNA sequences of Rickettsia spp. (spotted fever group) and Anaplasma spp. were detected in these ticks. Phylogenetic analyses revealed possible existence of undescribed Babesia spp. and Borrelia spp. This study illustrates potential threat to domestic animals and humans from tick-borne pathogens.

Objective To investigate the value of renal CT volumetric texture analysis with machine learning radiomics in assessment of pathological grade of clear cell renal cell carcinoma(ccRCC). Methods Thirty-four biopsy-confirmed ccRCC subjects who had four-phase CT scanning (NC:non-contrast, CM: Corticomedullary, N: Nephrographic, E: Excretory) were collected retrospectively from June 2013 to October 2017 for the study.Non-rigid registration was performed on multi-phase CT images in reference to CM-phase.Each lesion was segmented on CM-phase CT images using our in-house volumetric image analysis platform,"3DQI".A set of fifty-nine volumetric textures,including histogram,gradient,gray level co-occurrence matrix(GLCM),run-length(RL),moments,and shape,was calculated for each segment lesion in each phase as parameters for the training/testing of Random Forest (RF) classifier. Four groups according to pathological Fuhrman grade on a scaleⅠtoⅣ,these tumors were then divided into low(Ⅰ+Ⅱ) and high grade ( Ⅲ + Ⅳ) groups. Feature selection was performed by Boruta algorithm. A 10-fold cross-validation method was applied to validate the RF performance by receiver operating characteristic (ROC) curves analysis to determine the diagnostic accuracy of the model. Results Subjects were divided into four groups by Fuhrman grade on a scaleⅠtoⅣ:3 cases gradeⅠ,19 cases gradeⅡ,8 cases gradeⅢand 4 cases gradeⅣ.In CM-phase,kurtosis and long-run-emphasis(RLE)were selected the most important textures for ccRCC staging among 59 features. The area under curve (AUC) of ROC was 0.88 (79% sensitivity and 82% specificity)by using kurtosis and RLE textures.The mean values of kurtosis and RLE were(-20.00±22.00)×10-2and(3.00±0.40)×10-2for low group,whereas(31.00±32.00)×10-2and(5.00± 0.02)×10-2for high group.Within the mean±SD range of statistics,radiomics can distinguish between low and high grade tumors.In multi-phase analysis,three most important features were selected among 236(59× 4) textures: kurtosis (CM-phase), GLCM homogeneity I (HOMO 1) (E-phase), and GLCM homogeneity 2 (HOMO2)(E-phase).The mean values of HOMO 1(E-phase)and HOMO 2(E-phase)were(19.00±0.03)× 10-2and(11.00±0.02)×10-2for low group,whereas(22.00±0.03)×10-2and(14.00±0.02)×10-2for high group. The AUC was 0.92(93% sensitivity and 87% specificity)by using these three textures. Conclusion This study has demonstrated that renal CT volumetric texture analysis with machine learning radiomics could preoperative accurately perform cancer staging for ccRCC.

Chronic heart failure(CHF) is the end stage of various cardiovascular diseases and is one of the common diseases among the elderly in China. Sarcopenia is a geriatric syndrome characterized by the age-related loss of skeletal muscle quantity and/or mass and the decline in muscle strength and/or physical performance. There is a common pathophysiological mechanism of CHF and sarcopenia, and they interact with each other and influence each other′s prognosis. There are few confirmed effective treatments for sarcopenia, and even fewer measures that can help slow the progression of both syndromes. However, on the basis of heart failure treatment hormone therapy, exercise training and nutritional support may slow the progression of sacopenia. This article reviews the recent advances in the relationship between elderly CHF patients with sarcopenia and related treatment.

Objective To investigate the differences in efficacy and safety in the treatment of patients with cerebral palsy at different grades of the Gross Motor Function Classification System(GMFCS)by selective posterior rhizotomy(SPR).Methods Relevant literatures on SPR treatment for cerebral palsy were retrieved from Pubmed,Embase,Web of Science,China Biology Medicine disc,China National Knowledge Infrastructure(CNKI),Wanfang Database,and VIP Database.Clinical trials on SPR treatment for cerebral palsy were included for Meta-analysis.At least two re-searchers independently screened the literatures,extracted data,and assessed the quality of the liter-atures.Data analysis was performed by Review Manager 5.4 software.Results A total of 2,726 lit-eratures were retrieved,and 8 literatures were finally included after screening.The results of the Me-ta-analysis showed that the gross motor function and self-care ability of patients with cerebral palsy at all GMFCS grades improved significantly after surgery,and muscle tone decreased significantly after surgery(P＜0.05).In comparison of the improvement in gross motor function before and after SPR,patients with grades Ⅱ and Ⅲ of GMFCS benefited the most,followed by those with grade Ⅰ,and those with grades Ⅳ and Ⅴ benefited less.In terms of improving self-care ability,patients with grade Ⅰbenefited the most,followed by those with grade Ⅲ,and those with grades Ⅱ and Ⅳ benefited less.No significant adverse reactions were reported in previous literatures.Conclusion SPR is a relatively safe and effective treatment option for patients with cerebral palsy.Patients at grades Ⅱand Ⅲ of GMFCS benefit the most from SPR,and patients at grades Ⅳ and V with poor preoperative physical status can also benefit from SPR.

Objective To investigate the differences in efficacy and safety in the treatment of patients with cerebral palsy at different grades of the Gross Motor Function Classification System(GMFCS)by selective posterior rhizotomy(SPR).Methods Relevant literatures on SPR treatment for cerebral palsy were retrieved from Pubmed,Embase,Web of Science,China Biology Medicine disc,China National Knowledge Infrastructure(CNKI),Wanfang Database,and VIP Database.Clinical trials on SPR treatment for cerebral palsy were included for Meta-analysis.At least two re-searchers independently screened the literatures,extracted data,and assessed the quality of the liter-atures.Data analysis was performed by Review Manager 5.4 software.Results A total of 2,726 lit-eratures were retrieved,and 8 literatures were finally included after screening.The results of the Me-ta-analysis showed that the gross motor function and self-care ability of patients with cerebral palsy at all GMFCS grades improved significantly after surgery,and muscle tone decreased significantly after surgery(P＜0.05).In comparison of the improvement in gross motor function before and after SPR,patients with grades Ⅱ and Ⅲ of GMFCS benefited the most,followed by those with grade Ⅰ,and those with grades Ⅳ and Ⅴ benefited less.In terms of improving self-care ability,patients with grade Ⅰbenefited the most,followed by those with grade Ⅲ,and those with grades Ⅱ and Ⅳ benefited less.No significant adverse reactions were reported in previous literatures.Conclusion SPR is a relatively safe and effective treatment option for patients with cerebral palsy.Patients at grades Ⅱand Ⅲ of GMFCS benefit the most from SPR,and patients at grades Ⅳ and V with poor preoperative physical status can also benefit from SPR.

Objective:To investigate the effect of Fu-Fang-Yu-Jie(FFYJ)granules on LPS-induced inflammation model in bone marrow-derived macrophages(BMDM)and its intervention of FFYJ on nucleotide-bound oligomeric domain-like receptor protein 3(NLRP3)inflammatory signaling pathway in macrophages.Methods:Primary cells were extracted and isolated from the leg bone mar-row of C57BL/6 mice and BMDM macrophages were obtained after 7 days of induction with 50 ng/ml M-CSF.Groups included control group(Control),model group(LPS+ATP),FFYJ low dose group(FFYJ 50 μg/ml),FFYJ medium dose group(FFYJ 100 μg/ml),FFYJ high dose group(FFYJ 200 μg/ml)and positive drug dexamethasone group(DEX).BMDM in FFYJ treatment group and posi-tive drug group were pretreated for 1 hour before modeling.Lactate dehydrogenase(LDH)release assay was used to detect the level of LDH in the supernatant of each group of cells;ELISA was used to detect the level of inflammatory factors TNF-α,IL-6 and IL-1β in the supernatant of each group of cells;qRT-PCR was used to detect the levels of TNF-α,IL-6 and IL-1β in each group of cells;protein levels of NF-κB,p-NF-κB,NLRP3,Caspase-1 p45,Caspase-1 p20 and GSDMD-N in each group of cells were detected by Western blot;inverted fluorescence microscope was used to observe the cell pyroptosis of each group after Hoechst-PI staining.Results:Compared with control group,the levels of LDH,TNF-α,IL-6 and IL-1β in the supernatant of the model group were signifi-cantly higher(P<0.000 1);the mRNA levels of TNF-α,IL-6 and IL-1β in the model group were significantly higher(P<0.000 1);the protein levels of p-NF-κB,NLRP3,Caspase-1 p20 and GSDMD-N were significantly higher(P<0.05);and the number of PI-posi-tive cells was significantly higher(P<0.05).Compared with the model group,FFYJ and DEX significantly reduced the levels of LDH,TNF-α,IL-6 and IL-1β in the supernatant of BMDMs(P<0.05);down-regulated the mRNA levels of TNF-α,IL-6 and IL-1β in the cells(P<0.05);and inhibited the expressions of p-NF-κB,NLRP3,Caspase-1 p20 and GSDMD-N protein expressions(P<0.05);and significantly reduced the number of PI-positive cells(P<0.05).Conclusion:FFYJ exerts anti-inflammatory effects by inhibiting NLRP3 inflammasome activation in BMDM macrophages.

Objective To investigate the ultrasonography discrepancy of hepatic neuroendocrine neoplasm ( hNEN ) derived from different primary locations and pathological grades . Methods From August 2010 to April 2016 ,50 consecutive patients with 54 hNENs confirmed by pathology and imaging diagnosis underwent baseline ultrasound ( BUS ) and contrast-enhanced ultrasound ( CEUS ) in the First Affiliated Hospital of Sun Yat-sen University were enrolled in the retrospective study . On the basis of primary sites ,all hNENs were divided into three groups :pancreas ,gastrointestinal tract and other location groups . According to the pathological grading ,the hNENs were divided into hNET ( hepatic neuroendocrine tumor) group and hNEC ( hepatic neuroendocrine carcinoma) group . The imaging features of BUS and CEUS were retrospectively investigated and the ultrasonography discrepancy of hNEN derived from different primary focus and pathological grades were compared . Results ① Comparison of hNEN derived from different primary sites :hNEN derived from pancreas and gastrointestinal tract were smaller than hNEN derived from other sites[ ( 3 .8 ± 2 .6) cm vs ( 8 .7 ± 7 .0) cm , P = 0 .037 ;( 2 .9 ± 2 .1) cm vs ( 8 .7 ± 7 .0) cm , P = 0 .005] ,but the difference between the pancreas and gastrointestinal tract was not statistically significant ( P = 1 .0 ) . As for the pattern of CEUS enhancement ,hNEN derived from pancreas and gastrointestinal tract usually showed homogenous enhancement ( 13/19 ,15/23) while hNEN derived from other primary sites usually showed heterogeneous enhancement ( 10/13) ,and the difference was statistically significant ( P = 0 .025) . ② Comparison of hNET and hNEC :the distinction of enhancement level in the portal phase of contrast-enhanced ultrasound was statistically significant ( P = 0 .033) . All hNEC appeared hypo-enhancement ,while a small part of hNET ( 7/31 ) appeared iso-enhancement .More hNEC lesions showed heterogenous enhancemnt than hNET in the arterial phase ( 14/21 vs 9/31 , P = 0 .007) ,and there were less hNEC lesions presented tumor bleeding vessels than hNET ( 8/21 vs 22/31 , P = 0 .019 ) . Conclusions hNEN derived from pancreas and gastrointestinal tract are in a manner similar to hNEN derived from other primary sites on ultrasonography , so it' s difficult to identify in ultrasound . The ultrasonographic features of hNEC are more resemble to malignancy and it can be distinguished from hNET .

Objective:To analyze the EGFR mutation profile of lung cancer patients in Yunnan, and to provide evidence for clinical personalized treatment.Methods:Demographic and clinical data of 2 967 lung cancer patients undergoing EGFR identification were collected and analyzed from January 2014 to August 2019 in Yunnan Cancer Hospital.Results:The proportion of EGFR mutation in 2 967 patients with lung cancer was 46.2%. Univariate analysis showed that the proportion of EGFR mutation in women was higher than that in men ( P<0.001) and displayed a downward trend with age ( P=0.03). The mutation rate of ethnic minorities was higher than Han ( P=0.012). Mutation rate in patients without smoking history was higher than those with smoking history ( P<0.001), and patients without drinking history was higher than patients with drinking history ( P<0.001). Mutation rate in patients without family history of lung cancer was higher than those with family history ( P=0.008). The mutation rate of adenocarcinoma was higher than other pathological types ( P<0.001). The mutation rate was different among stages, and it was higher in early patients than that in advanced patients ( P<0.001). The mutation rate of tissue specimens was higher than those of cytology and peripheral blood samples ( P<0.001). The mutation rate of Xuanwei area was lower than that in non-Xuanwei area ( P<0.001). Multivariate analysis showed that gender ( P<0.001), age ( P=0.036), smoking history ( P<0.001), pathological type ( P<0.001), specimen type ( P<0.001), and whether or not Xuanwei area ( P<0.001) were the independent factors of EGFR mutation.The EGFR mutation was more common in female, non-smokers, adenocarcinoma, non-Xuanwei area, tissue specimen and young lung cancer patients.The mutation types of EGFR in 1 370 cases mainly included 19-Del and L858R. The predominant mutation of EGFR in Xuanwei area was L858R, while in non-Xuanwei area was 19-Del.The mutation rates of G719X, G719X+ L861Q, G719X+ S768I, and S768I in Xuanwei were higher while the mutation rates of 19-Del, L858R, and 20-ins were lower than non-Xuanwei area ( P<0.05). The 19-Del mutation rate of ethnic minorities is higher than that of Han ( P<0.001). The combined mutation rate of G719X, L861Q in Han was higher than that of ethnic minorities ( P=0.005). Conclusions:The EGFR mutation rate in lung cancer patients in Yunnan is similar to Asian and Chinese, and higher in female, non-smokers, adenocarcinomas, young and non-Xuanwei area patients. The most common types of EGFR mutation in Yunnan are 19-Del and L858R. The predominant mutation of EGFR in Xuanwei area is L858R, while in non-Xuanwei area is 19-Del. The mutation rates of G719X, G719X+ L861Q, G719X+ S768I and S768I are higher in Xuanwei patients than those in non-Xuanwei patients. The combined mutation rate of G719X and L861Q in Han nationality is higher than that of ethnic minorities.