Objective To investigate the effect of the serum of patients with hepatopulmonary syndrome on the myogenic differentiation,proliferation and migration of human lung fibroblasts.Methods The human lung fibroblasts were seeded in plates or flasks and randomly divided into 2 groups (n =31each) using a random number table:serum of patients with hepato-pulmonary syndrome group and serum of healthy volunteer group.The human lung fibroblasts were incubated in the DMEM culture medium containing 10％ serum of patients with hepatopulmonary syndrome or in the DMEM culture medium containing 10％ serum of healthy volunteers.At 24,48 and 72 h of incubation (T1-T3),the expression of smooth muscle-α-actin (SM-α-actin) and smooth muscle myosin heavy chain (SM-MHC) in human lung fibroblasts was determined by Western blot,the proliferation of the human lung fibroblasts was determined using 3H-TDR incorporation assay,and the migration of the human lung fibroblasts was determined by Transwell chamber assay.Results Compared with serum of healthy volunteer group,the expression of SM-α-actin and SM-MHC in human lung fibroblasts was significantly up-regulated at each time point,and the proliferation and migration of the cells were significantly enhanced at T2,3 in serum of patients with hepatopulmonary syndrome group (P＜0.05).Compared with the value at T1,the expression of SM-α-actin and SM-MHC in human lung fibroblasts was significantly up-regulated,and the proliferation and migration of the cells were significantly enhanced at T2,3in serum of patients with hepatopulmonary syndrome group (P＜0.05).Compared with the value at T2,the expression of SM-α-actin and SM-MHC in human lung fibroblasts was significantly up-regulated,and the proliferation and migration of the cells were significantly enhanced at T3 in serum of patients with hepatopulmonary syndrome group (P＜0.05).Conclusion The serum of patients with hepatopulmonary syndrome can promote the myogenic differentiation,proliferation and migration of human lung fibroblasts.

Objective To investigate the risk factors for hemorrhagic transformation (HT) and poor outcomes in patients with acute ischemic stroke after mechanical thrombectomy.Methods The patients with acute ischemic stroke received mechanical thrombectomy were enrolled retrospectively.The demography,vascular risk factors and other clinical data of the patents were collected.The modified Rankin scale (mRS) was used to evaluate the clinical outcomes at day 90.Good outcome was defined as mRS score 0-2.The patients were divided into either a HT group or a non-HT group according to their HT conditions.Multivariate logistic regression analysis was used to identify the independent risk factors for HT and poor outcomes.Results A total of 48 patients with acute ischemic stroke received mechanical thrombectomy were enrolled,including 25 males (52.1％).Their mean age was 64.77± 9.14 years.The mean National Institutes of Health Stroke Scale (NIHSS) score was 17.70 ± 3.77.Twenty-two patients (45.8％) occured HT,of which 9 were symptomatic HT;24 (50.0％) had good outcomes.The proportion of males in the HT group was significantly lower than that in the non-HT goup (30.4％vs.72.0％;x2 =8.293,P =0.004),while the proportions in patients with diabetes (65.2％ vs.36.0％;x2 =4.090,P =0.043) and atrial fibrillation (78.3％ vs.44.0％;x2 =5.880,P =0.015),as well as the baseline fasting blood glucose level (8.514 ± 4.400 mmol/L vs.6.354 ± 1.472 mmol/L;t =2.319,P =0.025) were significantly higher than those in the non-HT group.Multivariate logistic regression analysis showed that the atrial fibrillation (odds ratio [OR] 6.136,95％ confidence interval [CI] 1.617-23.291;P =0.042) was a risk factor for the occurrence of HT after mechanical thrombectomy.The proportion of diabetic patients (29.2％ vs.70.8％;x2 =8.333,P=0.04) and baseline NIHSS score (16.050±4.865 vs.19.210±4.423);t=2.310,P=0.026) of the good outcome group were significantly lower than those of the poor outcome group,while the proportions of patients in atrial fibrillation (75.0％ vs.45.8％;x2 =4.269,P =0.039),anterior circulation stroke (87.5％ vs.62.5％;x2 =4.000,P =0.046) middle cerebral artery (75.0％ vs.29.2％;x2 =10.113,P =0.006),vertebral basilar artery (37.5％ vs.12.5％;x2 =10.113,P =0.006) occlusion and parenchymal hematoma (33.3％ vs.4.1％;P=0.011) were significantly higher than the poor outcome group.Multivariate logistic regression analysis showed that diabetes (OR 5.898,95％ CI 1.699-20.479;P=0.005),baseline NIHSS score (OR 1.167,95％ CI 1.011-1.347;P =0.035),and parenchymal hematoma (OR 1.295,95％ CI 1.099-1.875;P=0.028) were the independent risk factors for poor outcomes.Conclusions Atrial fibrillation is an independent predictor of HT risk in patients with acute ischemic stroke after mechanical thrombectomy.Diabetes mellitus,higher baseline NIHSS score,and concurrent brain parenchymal hematoma are the independent predictors of poor outcomes.Therefore,the risk of HT and adverse outcomes should be fully assessed before mechanical thrombectomy in patients with acute ischemic stroke.

Objective To study the possibility of direct identification of pathogens from positive blood cul-tures by methods of separation gel tube -centrifugation.Methods 216 cases of positive blood culture were collected from 2015.7 to 2015.12.The bacterias were purified from blood culture bottle by separation gel tube.After washing 2 times,identified by MALDI -TOF MS.At the same time,traditional culture,smears and identification were done. Compared the results of identification by two methods.Results 216 cases of positive blood culture were single bacte-rial infection.By Gram stain,89 strains were Gram positive,119 strains were Gram negative and 8 strains were fungal spores.190 cases of positive blood culture were identified by MALDI -TOF MS,it concluded 67 Gram positive strains,111 Gram negative strains,4 anaerobe strains and 8 fungus.Compared with traditional culture,the coincidence rate reached up to 87.9%,Gram positive strains 78.8%,Gram negative strains 93.2%,anaerobe strains 100.0%and fungus 100.0%.Conclusion It takes less than 30 minutes purified from blood culture bottle by separation gel tube.And the time of identification is shorter than traditional culture.This method is good for clinical diagnosis and treatment.

With increasing use of carbapenem antibiotics , carbapenems-resistant gram-negative bacteria are spreading, and carbapenemase-producing is the main mechanism of carbapenems resistance . Rapid and accurate identification of carbapenemase and its type is of great importance to timely and effective treatment and control of infections .Chromogenic /Fluorogenic culture media, modified Hodge test and double disk synergy test are traditional methods for carbapenemase detection , but all are time-consuming. Biochemical method is more time efficient and with high sensitivity and specificity , but cannot be used to identify subtypes.Now matrix-assisted laser desorption ionization -time-of-flight mass spectrometry (MALDI-TOF MS) has been successfully applied in the identification of species , subtypes and detection of drug -resistant genes.And among various carbapenemase gene detection techniques , next generation sequencing (NGS) can also be used for the detection of integrons , transposons and plasmids, which is important in both epidemiology and resistant mechanism studies .This article reviews the advantages and disadvantages of various methods for phenotype and gene detection of carbapenemase .

Objective To calculate the nonlinear features of motion in patients with chronic vestibular syndrome(CVS)using the largest Lyapunov exponent(LLE),and to verify the classification model's validity through machine learning algorithms.Methods A three-dimensional(3D)motion capture system was used to capture the joint motion trajectories of the subjects,which were determined using the LLE.The features of the chaotic trajectories were calculated as the input,and seven classifiers,namely the ID3 decision tree,Adaboost,C45 decision tree,Bayesian classification,Naive Bayes,and support vector machine,were used for classification.Results A total of 17 sets of trajectories from 16 joints were in the chaotic state,and the average energy,enhanced wavelength,and kurtosis of the motion trajectories in the experimental group showed significant differences(P<0.05).The ID3 decision tree classifier showed optimal performance with 100%prediction accuracy,recall,and F1-score.Conclusions Chaotic features may contain high personality differences in patients with CVS and can improve the accuracy of machine learning algorithms for recognition.These findings provide a reference for early identification and motor rehabilitation of patients with CVS.

Objective To evaluate the role of microglia in paraventricular nucleus (PVN) in sus-ceptibility to depression in rats with chronic visceral pain. Methods Ninety-six pathogen-free healthy male Sprague-Dawley rats, aged 8 days, were divided into 6 groups (n= 16 each) using a random number table: sham operation group (S group), chronic visceral pain group (CHVP group), sham operation plus colorectal distension group (S+C group), chronic visceral pain plus colorectal distension group (CHVP+C group), chronic visceral pain plus phosphate buffer solution plus colorectal distension group (CHVP+P+C group) and chronic visceral pain plus minocycline plus colorectal distension group (CHVP+M+C group). Colorectal distension was not performed in S group. In CHVP group, chronic visceral pain was induced by performing colorectal distension twice daily on postnatal days 8, 10, and 12. Phosphate buffer solution 0. 5μl was injected into PVN by stereotaxic method at 8th week after birth in CHVP+P+C group, and 2％ mi-nocycline 0. 5 μl was injected into PVN at 8th week after birth in CHVP+M+C group. Eight rats in each group were selected 2 h later for measurement of visceral pain threshold. In S+C, CHVP+C, CHVP+P+C and CHVP+M+C groups, colorectal distension was performed for 2 times, open field test and sucrose preference test were then performed, the rats were sacrificed and PVN was removed for determination of micro-glial activation by immunofluorescence. Results The pain threshold was significantly decreased in CHVP, CHVP+C, CHVP+P+C and CHVP+M+C groups as compared with S and S+C groups (P<0. 05). The pain threshold was significantly increased in CHVP+M+C group when compared with CHVP +P +C group (P<0. 05). Compared with S, CHVP and S+C groups, the total locomotor distance, the number of rea-ring and sucrose consumption were significantly reduced, and the proportion of activated microglia in PVN was increased in CHVP+C, CHVP+P+C and CHVP+M+C groups (P<0. 05). Compared with CHVP+P+C group, the total locomotor distance, the number of rearing and sucrose consumption were significantly in-creased, and the proportion of activated microglia in PVN was decreased in CHVP+M+C group (P<0. 05). Conclusion Microglia in PVN is involved in regulation of susceptibility to depression in rats with chronic visceral pain.

The early diagnosis of primary hepatocellular carcinoma combined with portal vein tumor thrombus is very difficult,and when the patients are diagnosed,they are usually at an advanced stage,and the optimal treatment time has missed. In recent years, more and more scholars believe that the diagnosis and treatment of hepatocellular carcinoma with portal vein tumor thrombus need multi?disciplinary diagnosis and treatment,it also has a certain effect in the clinic, it is expected to make the early diagnosis and the early treatment for patients with hepatocellular carcinoma combined with portal vein tumor thrombus in the future,and form a standardized treatment process.

Objective: To compare droplet digital PCR (ddPCR) and Super-amplification refractory mutation system (ARMS) in the detection of T790M mutation of epidermal growth factor receptor (EGFR) in the plasma of non-small cell lung cancer patients who had developed resistance to EGFR tyrosine kinase inhibitor (TKI) , and to investigate the clinical application of ddPCR. Methods: Plasma samples were collected from non-small cell lung cancer patients who had acquired EGFR-TKI resistance at Shanghai Pulmonary Hospital, Tongji University, from May 2017 to November 2017. Extracted ctDNA was analyzed by ddPCR and Super-ARMS to evaluate the T790M mutation status of EGFR gene. Results: A total of 37 patients with activating EGFR mutation that acquired resistance to EGFR-TKI were selected in the study, including 17 male and 20 female with a median age of 64 years (range 40-83 years). Before TKI treatment, all the patients harbored EGFR inhibitor sensitive mutations but without T790M mutation. After acquiring resistance to EGFR-TKI treatment, the T790M mutation rate detectable by ddPCR was 45.9% (17/37). In contrast, the mutation rate of T790M detectable by Super-ARMS was 35.1% (13/37, P<0.05). For the 13 positive cases detected by Super-ARMS (ΔCt<8), they were all positive by ddPCR assay; Among the 10 negative cases detected by Super-ARMS (ΔCt≥8), there were 3 cases positive by ddPCR assay. For patients without ΔCt by Super-ARMS assay, there was one weak positive case detectable by ddPCR assay. Among 17 EGFR T790M positive patients, 9 received EGFR inhibitor Osimertinib treatment, and 7 of them had good therapeutic response after the treatment. Conclusions While a significant correlation between the two methods is shown. ddPCR is more sensitive than Super-ARMS in the detection of EGFR T790M mutation, indicating that it is a better method in guiding target drug therapy of non-small cell lung cancer patients after acquiring the resistance to EGFR-TKI.

Objective To evaluate the efficacy of stroke volume variation (SVV) combined with controlled low central venous pressure (CLCVP)-directed fluid therapy in the patients undergoing liver cancer resection.Methods Seventy American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients,aged 40-60 yr,with body mass index of 20-25 kg/m2,scheduled for elective liver cancer resection under general anesthesia,were divided into 2 groups (n=35 each) using a random number table:routine fluid replacement group (R group) and SVV combined with CLCVP-guided fluid replacement group (SC group).In R group,routine fluid replacement included compensatory volume expansion,physiological requirement,cumulative loss,continued loss (intraoperative blood loss) and 3rd space losses,maintaining mean arterial pressure＞70 mmHg,central venous pressure＜4 cmH2O and heart rate ＜ 100 bpm.Central venous pressure was maintained ＜4 cmH2O and SVV ＜ 12％ during operation in SC group.The operation time,total amount of crystalloid and colloid solution infused,urine volume and development of intraoperative hypotension and bradycardia were recorded.Blood samples from the left radial artery and central vein were collected before anesthesia induction and at the end of operation for measurement of the blood lactate concentration,and the oxygen supply index,oxygen consumption index and oxygen uptake rate were calculated.Blood samples from the ulnar vein were collected before anesthesia induction and at the end of operation for determination of serum β2-microglobulin concentrations.The development of intraoperative adverse cardiovascular events was recorded,and the occurrence of postoperative complications was observed.Results Compared with R group,the total amount of crystalloid solution infused was significantly decreased,the total amount of colloid solution infused and urine volume were increased,the incidence of intraoperative hypotension and bradycardia was decreased,oxygen supply index,oxygen consumption index and oxygen uptake rate were increased at the end of operation,and the lactate concentration in arterial blood,serum β2-microglobulin concentration and rate of postoperative pulmonary infection were decreased in SC group (P＜0.05).Conclusion SVV combined with CLCVP-directed fluid therapy produces better efficacy than routine fluid replacement in the patients undergoing liver resection.