Effect of Linggui Zhugantang on Mitochondrial Fusion-fission and Sirt3/ AMPK Signaling Pathway in Chronic Heart Failure Rats After Myocardial Infarction / 中国实验方剂学杂志

ObjectiveTo investigate the effects of Linggui Zhugantang on mitochondrial fission and fusion and silencing information regulator 3（Sirt3）/adenosine monophosphate dependent protein kinase （AMPK） signaling pathway in chronic heart failure （CHF） rats after myocardial infarction （MI）. MethodSD rats randomly divide into sham operation group （normal saline ，thread only without ligature）， model group （normal saline， ligation of the left anterior descending coronary artery proximal to the heart）， Linggui Zhugantang group （4.8 g·kg-1） and Captopril group （0.002 57 g·kg-1）， with 10 rats in each group. Administere drug continuously for 28 days. Echocardiography detected cardiac function parameters. Hematoxylin eosin （HE） staining observed the pathological changes of the heart. Immunofluorescence detected the levels of reactive oxygen species （ROS）. JC-1 detect mitochondrial membrane potential. Colorimetry measure adenosine triphosphate （ATP）， superoxide dismutase （SOD）， malondialdehyde （MDA）， mitochondrial respiratory chain complex activity （Ⅰ-Ⅳ）. TdT-mediated dUTP nick end labeling （TUNEL） staining detected the apoptosis rate of myocardial tissue. Western blot detected protein expression levels of Sirt3， phosphorylated AMPK （p-AMPK）， phosphorylated dynamic-related protein 1（p-Drp1）， mitochondrial fission protein 1（Fis1）， mitochondrial fission factor （MFF）， optic atrophy protein 1（OPA1）. ResultCompared to the sham group， the left ventricular end diastolic diameter （LVIDd） and left ventricular end systolic diameter （LVIDs） were significantly increased in model group （P<0.01）， while the left ventricular short axis shortening rate （LVFS） and left ventricular ejection fraction （LVEF） were significantly decreased （P<0.01）. There were inflammatory cell infiltration and obvious pathological injury in myocardial tissue. ROS， MDA levels and myocardial cell apoptosis rate were significantly increased （P<0.01）， SOD level， ATP content， and membrane potential were significantly decreased （P<0.01）. The activity of mitochondrial respiratory chain complexes （Ⅰ-Ⅳ） was significantly decreased （P<0.01）. Levels of p-Drp1， Fis1， MFF proteins were significantly up-regulated （P<0.01）， while Sirt3， p-AMPK， OPA1 proteins level were significantly down-regulated （P<0.01）. Compared with model group， LVIDd and LVIDs were significantly decreased （P<0.01）， LVEF and LVFS were significantly increased （P<0.01）. Inflammatory cell infiltration and pathological damage of myocardial tissue were significantly relieved. ROS， MDA levels and myocardial cell apoptosis rate were significantly decreased in Linggui Zhugantang group and Captopril group （P<0.01）， SOD level， ATP content， and membrane potential significantly increased （P<0.01）. The activity of mitochondrial respiratory chain complexes （Ⅰ-Ⅳ） increased significantly （P<0.01），and p-Drp1， Fis1， MFF protein levels were significantly down-regulated （P<0.01）， Sirt3， p-AMPK， OPA1 protein were significantly up-regulated （P<0.01）. ConclusionLinggui Zhugantang can alleviate oxidative stress and apoptosis damage of myocardial cells， maintain mitochondrial function stability， and its effect may be related to mitochondrial mitosis fusion and Sirt3/AMPK signaling pathway.

Relationship between blood-brain barrier changes and drug metabolism under high-altitude hypoxia: obstacle or opportunity for drug transport?

The blood-brain barrier is essential for maintaining the stability of the central nervous system and is also crucial for regulating drug metabolism, changes of blood-brain barrier's structure and function can influence how drugs are delivered to the brain. In high-altitude hypoxia, the central nervous system's function is drastically altered, which can cause disease and modify the metabolism of drugs in vivo. Changes in the structure and function of the blood-brain barrier and the transport of the drug across the blood-brain barrier under high-altitude hypoxia, are regulated by changes in brain microvascular endothelial cells, astrocytes, and pericytes, either regulated by drug metabolism factors such as drug transporters and drug-metabolizing enzymes. This article aims to review the effects of high-altitude hypoxia on the structure and function of the blood-brain barrier as well as the effects of changes in the blood-brain barrier on drug metabolism. We also hypothesized and explore the regulation and potential mechanisms of the blood-brain barrier and associated pathways, such as transcription factors, inflammatory factors, and nuclear receptors, in regulating drug transport under high-altitude hypoxia.

A Real-world Study on the Incidence and Outcome of Immune-related Adverse Events in Lung Cancer Patients / 中国肺癌杂志

BACKGROUND@#Immune-related adverse events (irAEs) are commonly occurred in patients treated with immune checkpoint inhibitors. However, evidence of irAEs derived from the Chinese population is relatively lacking. The aim of this study was to investigate the incidence and outcomes of irAEs in Chinese patients with lung cancer after receiving immune checkpoint inhibitors (ICIs).@*METHODS@#Clinical and follow-up data from lung cancer patients who received at least one time of ICIs from January 2018 to September 2021 at Huadong Hospital, Fudan University were included. Statistical descriptions and Kaplan-Meier method were used to analyze the overall incidence of irAEs, as well as the incidence and outcomes of each type of irAEs.@*RESULTS@#135 patients were included in the study. 106 patients (78.5%) presented at least one type of irAEs, and the median time to first irAEs onset was 28 d. Most irAEs occurred at early time after treatment, and most irAEs were mild-moderate and reversible. 57 patients (42.2%) died at the study cutoff. The mortality rate of severe irAEs was 12.6% (n=17), and among them 7 patients (41.2%) died of pneumonitis. The median progression-free survival (PFS) and overall survival (OS) time of the total population was 505 d (95%CI: 352-658) and 625 d (95%CI: 491-759), respectively. Patients who presented any irAEs achieved a longer PFS than those who did not (median PFS: 533 d vs 179 d, P=0.037, HR=0.57), while patients who presented skin toxicities achieved a longer OS than patients who did not (median OS: 797 d vs 469 d, P=0.006, HR=0.70).@*CONCLUSIONS@#In real-world settings, irAEs in lung cancer patients were commonly observed, with pneumonitis as the most common fatal irAEs. In addition, patients who presented any irAEs may tend to achieve a longer PFS.

Protective Effect of Linggui Zhugantang Medicated Serum Against H2O2-induced Injury in H9c2 Cells by Regulating PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of Linggui Zhugantang （LGZGT）-medicated serum against H2O2-induced injury in H9c2 cells and its relationship with the phosphatidylinositol 3- kinase/protein kinase B （PI3K/Akt） signaling pathway. MethodThe LGZGT-medicated serum and blank serum were prepared based on serum pharmacology. H9c2 cells were cultured in vitro and divided into a normal group， an H2O2 group， a 20% blank serum group， and a 20% LGZGT-medicated serum group. The cells were treated with corresponding drugs for 12 h and cultured with 100 μmol·L-1 H2O2 for another 6 h. The effect of 20% LGZGT-medicated serum on the proliferation activity of H9c2 cells induced by H2O2 was detected by cell counting kit-8 （CCK-8） assay. Mitochondrial reactive oxygen species （ROS） level was detected by the fluorescence probe. The levels of malondialdehyde （MDA）， lactate dehydrogenase （LDH）， catalase （CAT）， and glutathione peroxidase （GSH-Px） were detected by colorimetry. Western blot was used to detect the protein expression levels of phosphoinositide 3-kinase （PI3K）， phosphorylated-PI3K （p-PI3K）， protein kinase B （Akt）， and phosphorylated-Akt （p-Akt）. Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） was used to detect mRNA expression of PI3K and Akt. Flow cytometry was used to detect the apoptosis rate. After the addition of PI3K inhibitor LY294002， the levels of mitochondrial ROS， LDH， and GSH-Px， protein expression of PI3K， p-PI3K， Akt， and p-Akt， and cell apoptosis rate were detected. ResultCompared with the normal group， the H2O2 group showed blunted cell viability （P<0.01）， increased levels of mitochondrial ROS， MDA， and LDH （P<0.01）， decreased levels of CAT and GSH-Px （P<0.01）， reduced phosphorylation and mRNA expression of PI3K and Akt （P<0.05， P<0.01）， and increased apoptosis rate （P<0.01）. Compared with the H2O2 group， the 20% LGZGT-medicated serum group showed potentiated cell viability， reduced levels of mitochondrial ROS， MDA， and LDH （P<0.01）， increased levels of CAT and GSH-Px （P<0.01）， up-regulated phosphorylation and mRNA expression of PI3K and Akt （P<0.05， P<0.01）， and decreased apoptosis rate （P<0.01）. The combined use of LGZGT-medicated serum and inhibitor LY294002 reversed the above-mentioned effects of LGZGT-medicated serum on H9c2 cells （P<0.05， P<0.01）. ConclusionThe protective effect of LGZGT-medicated serum on H2O2-induced H9c2 cell injury may be related to the regulation of the PI3K/Akt signaling pathway to reduce oxidative stress and apoptosis.

A Decade's Review of miRNA: A Center of Transcriptional Regulation of Drugmetabolizing Enzymes and Transporters under Hypoxia.

BACKGROUND: Hypoxia has a negative effect on the cardiovascular system, nervous system, and metabolism, which contributes to potential changes in drug absorption, distribution, metabolism, and excretion (ADME). However, hypoxia can also alter the expression of microRNA (miRNA), thereby regulating drug-metabolizing enzymes, transporters, and ADME genes, such as hypoxia-inducible factor, inflammatory cytokine, nuclear receptor, etc. Therefore, it is crucial to study the role of miRNA in the regulation of drug-metabolizing enzymes and transporters under hypoxia. METHODS: A systematic review of published studies was carried out to investigate the role of miRNA in the regulation of drug-metabolizing enzymes and transporters under hypoxia. Data and information on expression changes in miRNA, drug-metabolizing enzymes, and transporters under hypoxia were analyzed and summarized. RESULTS: Hypoxia can up or down-regulate the expression of miRNA. The effect of hypoxia on Cytochrome P450 (CYP450) is still a subject of debate. The widespread belief is that hypoxia decreased the activity and expression of CYP1A1, CYP1A2, CYP2E1, and CYP3A1 and increased those of CYP3A6 and CYP2D1 in rats. Hypoxia increased the expression of a multidrug resistance-associated protein, breast cancer resistance protein, peptide transporter, organic cation transporter, and organic anion transporter. miRNA negatively regulated the expression of drugmetabolizing enzymes and transporters. CONCLUSION: The findings of this review indicated that miRNA plays a key role in the expression changes of drugmetabolizing enzymes and transporters under hypoxia.

Evaluation of the performance of systems for whole blood C-reactive protein detection: a multi-center study / 中华检验医学杂志

Objective:To explore the performance of the commonly used whole blood C-reactive protein (CRP) detection systems and give related recommendation on the performance requirements of detection systems.Methods:A total of 7 540 venous blood samples from 26 maternal, child and children′s hospitals were collected to conduct this multi-center study on the analytical performance of 5 commonly used whole blood CRP detection systems from March to April in 2019. The blank check, carryover, repeatability, intermediate precision, linearity, sample stability, influence of hematocrit/triglyceride/bilirubin, comparison with SIEMENS specific protein analyzer and trueness were evaluated. The 5 systems included BC-5390CRP autohematology analyzer, AstepPLUS specific protein analyzer, Ottoman-1000 Automated Specific Protein POCT Workstation, i-CHROMA Immunofluorometer equipment Reader and Orion QuikRead go detecting instrument. The 5 systems were labeled as a, b, c, d and e randomly.Results:Within the 5 systems, all values of blank check were less than 1.00 mg/L, the carryovers were lower than 1.00%. The repeatability of different ranges of CRP concentrations including 3.00-10.00, 10.00-30.00 and>30.00 mg/L were less than 10.00%, 6.00% and 5.00%, respectively, and the intermediate precision was less than 10.00%. The linearity correlation coefficients of the 5 systems were all above 0.975, while the slope was within 0.950-1.050. Whole blood samples were stable within 72 hours both at room temperature (18-25 ℃) and refrigerated temperature (2-8 ℃). The CRP results were rarely influenced by high triglyceride or bilirubin, except for the immmunoturbidimetric test based on microparticles coated with anti-human CRP F(ab) 2 fragments. When triglyceride was less than 15.46 mmol/L, the deviation of CRP was less than 10.00%. When bilirubin was less than 345.47 μmol/L, the deviation of CRP was less than 10.00%. CRP was more susceptible to Hct on the systems without Hct correction. The deviation of CRP between different Hct dilution concentration and 40% dilution concentration can reach as high as 67.48%. The correlation coefficients ( r) of 5 systems were all more than 0.975 in the range of 0-300.00 mg/L compared with Siemens specific protein analyzer. All systems passed the trueness verification using the samples with specified values of 12.89 and 30.60 mg/L. Conclusion:The performance of 5 systems can basically meet the clinical needs, but it is suggested that the whole blood CRP detection system without automatic Hct correction should be modified manually.

An Observational Study of COVID-19 from A Large Healthcare System in Northern New Jersey: Diagnosis, Clinical Characteristics, and Outcomes

BackgroundNew Jersey was an early epicenter for the COVID-19 pandemic in the United States, yet information on hospitalized COVID-19 patients from this area is scarce. This study aimed to provide data on demographics and clinical features of a hospitalized patient population who were confirmed with infection by our in-house (CDI) real-time reverse-transcription polymerase chain reaction (RT-PCR) test. MethodsWe included consecutive patients who were admitted to Hackensack Meridian Health system hospitals with laboratory-confirmed diagnoses of COVID-19 at Hackensack University Medical Center by the CDI virus test between March 12, 2020, and April 8, 2020. Clinical data and viral testing results were collected and analyzed for characteristics associated with outcomes, as well as the correlation with viral load. ResultsA total of 722 patients were included in the study, with a median age of 63 (interquartile range (IQR), 51-75) and 272 (37.7%) females. Mortality of this case series was 25.8%, with a statistically significant linear increase observed from age 40 to [≥]80 by 10-year intervals. Viral load, as indicated by the cycle of threshold (Ct) values from the RT-PCR test, was significantly higher in the oldest patient group ([≥]80), and inversely correlated with survival. ConclusionsThis is the first report to describe the clinical characteristics and outcomes in a large hospitalized COVID-19 patient series from New Jersey. Findings from this study are valuable to the ongoing response of both nationwide healthcare networks and the medical research community. SummaryWe describe the diagnosis, clinical characteristics, and outcomes of a large hospitalized patient population in northern New Jersey during the early stages of the COVID-19 pandemic.

A descriptive study of the impact of diseases control and prevention on the epidemics dynamics and clinical features of SARS-CoV-2 outbreak in Shanghai, lessons learned for metropolis epidemics prevention

ObjectiveTo describe and evaluate the impact of diseases control and prevention on epidemics dynamics and clinical features of SARS-CoV-2 outbreak in Shanghai. DesignA retrospective descriptive study SettingChina ParticipantsEpidemiology information was collected from publicly accessible database. 265 patients admitted to Shanghai Public Health Center with confirmed COVID-19 were enrolled for clinical features analysis. Main outcome measurePrevention and control measures taken by Shanghai government, epidemiological, demographic, clinical, laboratory and radiology data were collected. Weibull distribution, Chi-square test, Fishers exact test, t test or Mann-Whitney U test were used in statistical analysis. ResultsCOVID-19 transmission rate within Shanghai had reduced over 99% than previous speculated, and the exponential growth has been stopped so far. Epidemic was characterized by the first stage mainly composed of imported cases and the second stage where >50% of cases were local. The incubation period was 6.4 (95% CI 5.3 to 7.6) days and the mean onset-admission interval was 5.5 days (95% CI, 5.1 to 5.9). Median time for COVID-19 progressed to severe diseases were 8.5 days (IQR: 4.8-11.0 days). By February 11th, proportion of patients being mild, moderate, severe and critically ill were 1.9%(5/265), 89.8%(238/265), 3.8%(10/265), 4.5%(12/265), respectively; 47 people in our cohort were discharged, and 1 patient died. ConclusionStrict controlling of the transmission rate at the early stage of an epidemic in metropolis can quickly prohibit the spread of the diseases. Controlling local clusters is the key to prevent outbreaks from imported cases. Most COVID-19 severe cases progressed within 14 days of disease onset. Multiple systemic laboratory abnormalities had been observed before significant respiratory dysfunction.

Regulatory effects of sphingosine kinase-2 on astrocyte function and EAE progression in mouse model / 中华微生物学和免疫学杂志

Objective:To investigate the regulatory effects of sphingosine kinase-2 (SphK2) on the function of activated astrocytes and the progression of experimental autoimmune encephalomyelitis (EAE) in mice.Methods:Primary mouse astrocytes were isolated from wild-type (WT) C57BL/6 mice and sphk2 gene knock-out ( sphk2 -/-) mice and stimulated in vitro with interleukin 17 (IL-17). Real-time PCR was used to measure the expression of inflammatory cytokines and chemokines at mRNA levels. Western blot and immunofluorescence were used to detect the expression of glial fibrillary acidic protein (GFAP) and the phosphorylation of signal transducer and activator of transcription 3 (p-STAT3). An EAE mouse model was constructed using myelin oligodendrocyte glycoprotein 35-55 (MOG 35-55) polypeptide. Western blot was used to detect the expression of GFAP and p-STAT3 at protein level and real-time PCR was used to detect the expression of inflammatory cytokines and chemokines at mRNA level in spinal cords. Hematoxylin-Eosin (HE) and Luxol fast blue (LFB) staining were used to observe the changes in inflammatory cell infiltration and demyelination in spinal cords. Results:Compared with the WT group, the phosphorylation of STAT3 was obviously reduced in in vitro activated mouse astrocytes of sphk2 -/- mice, and the expression of inflammatory cytokines and chemokines including monocyte chemotactic protein 1 (MCP-1), TNF-α and IL-6 at mRNA level was also significantly decreased. Compared with the WT EAE group, changes in the above-mentioned cytokines and relative proteins in sphk2 -/- EAE mice in vivo were similar to those in vitro. Moreover, inflammatory cell infiltration and demyelination were significantly reduced in spinal cords of sphk2 -/- EAE mice. However, no significant difference in in vitro or in vivo GFAP expression was observed between WT and sphk2 -/- mice. Conclusions:SphK2 might regulate the function of reactive astrocytes through STAT3 molecular pathway, thereby regulating the production of inflammatory cytokines and chemokines and participating in the pathological process of EAE.

Effect of X-ray irradiation on pharmacokinetics of irinotecan hydrochloride and expression of CES1 and CYP3A1 in rats.

Concurrent chemoradiation with irinotecan hydrochloride (CPT-11) is accepted for cancer treatment. However, the effects of X-ray irradiation on chemotherapeutics in the plasma remain unclear. We evaluated the pharmacokinetics of CPT-11 in rats after exposure to X-ray irradiation and examined the changes of protein and mRNA expression of CES1 and CYP3A1. The X-ray irradiation with 1 Gy and 5 Gy was delivered to the whole body of rats. CPT-11 at 30 and 60 mg/kg, respectively, was intravenously infused 24 h after irradiation. CPT-11 was determined by RP-HPLC in plasma. ELISA and PCR were used to analyze the protein and mRNA expression of CES1 and CYP3A1, respectively. Compared with control rats, the X-ray irradiation decreased the AUC of CPT-11 (30 mg/kg) by 15.6% at 1 Gy and 39.0% at 5 Gy and increased the CL by 60.0% at 5 Gy. The X-ray irradiation could also decrease the AUC of CPT-11 (60 mg/kg) and increase the CL. In addition, the protein and mRNA expression of CES1 and CYP3A1 were increased significantly in rats after irradiation. This study found significant changes in the pharmacokinetics of CPT-11 in rats after exposure to X-ray irradiation, and they might be due to significant increases in the expressions of CYP3A1 and CES1. The pharmacokinetics of CPT-11 should be rechecked, and the optimal CPT-11 dose should be reevaluated during concurrent chemoradiation therapy.

Simultaneous Determination of Contents of 8 Components in Eucommia ulmoides Leaves by HPLC / 中国药房

OBJECTIVE： To establish content determination method for simultaneously determining aucubin，geniposidic acid，catechin，chlorogenic acid，asperuloside，rutin，isoquercitrin and astragalin in Eucommia ulmoides leaves. METHODS：HPLC method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with the mobile phase consisted of acetonitrile-0.1％ phosphoric acid（gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 203 nm for aucubin and catechin，239 nm for geniposidic acid and asperuloside，220 nm for chlorogenic acid，354 nm for rutin and isoquercitrin，and 266 nm for astragalin. The sample size was 5 μL. RESULTS： The linear range of aucubin， geniposidic acid， catechin， chlorogenic acid， asperuloside， rutin， isoquercitrin and astragalin were 0.812-6.090 μg（r＝0.999 3），0.438-3.285 μg（r＝0.999 2），0.045-0.336 μg（r＝0.999 2），0.882-6.615 μg（r＝0.999 3），0.097-0.726 μg（r＝0.999 1），0.064-0.483 μg（r＝0.999 3），0.048-0.360 μg（r＝0.999 1） and 0.014-0.108 μg（r＝0.999 7），respectively. RSDs of precision， stability and reproducibility tests were all lower than 3.5％（n＝6）. The average recovery rates were 101.60％，103.06％，99.77％，96.93％，98.17％，96.75％，98.97％ and 99.60％，with RSDs of 1.42％，2.65％，2.78％，2.05％，2.26％，0.93％，2.79％ and 3.08％，respectively（n＝6）. The contents of aucubin， geniposide， catechin， chlorogenic acid， plantain， rutin， isoquercetin and astragaloside in 12 batches of E. ulmoides leaves from different collection time and planting varieties were 10.903-17.245， 5.578-7.892， 0.198-0.440， 13.890-19.782， 1.008-1.547， 1.102-2.396， 0.267-0.701， 0.150-0.412 mg/g， respectively. The content fluctuated greatly. The contents of aucubin， geniposide， catechin， chlorogenic acid， rutin and pinoresinol diglucoside in cortex of E. ulmoides were 0.299， 0.123， 0.580， 0.112， 0.026，1.961 mg/g， respectively. CONCLUSIONS： The method is simple， reproducible and accurate. It can be used to evaluate the quality of E. ulmoides leaves. There are obvious differences in composition and content of components in different medicinal parts （cortex， leaves） of E. ulmoides.

Preliminary evaluation of reducing residual risk of blood transfusion-transmitted infection by using HBV DNA detection / 中华实验和临床病毒学杂志

Objective@#To evaluate the reduction of the residual risk of blood transfusion- transmitted hepatitis B virus (HBV), using nucleic acid detection(NAT)test for enzyme linked immunosorbent assay (ELISA) qualified volunteer-donor bloods in Quzhou area after NAT was developed.@*Methods@#Specimens were collected from March 2016 to March 2017, detected by ELISA twice with two different reagents and NAT only once. The residual risks of blood transfusion-transmitted HBV infection were calculated by mathematical model of risk evaluation.@*Results@#Totally 27 646 specimens were collected from March 2016 to March 2017, which included 76 specimens that were both ELISA and NAT positive, 31 specimens were ELISA negative but NAT positive.The total number of NAT positive specimens was 107.The residual risk of HBV by ELISA test was 28.2×10 -5and NAT test was 13.0×10-5.@*Conclusions@#NAT detection can greatly reduce the residual risk of blood transfusion-transmitted HBV infection, and provide effective value for bloods safety in practice.

Induced pluripotent stem cells differentiate into intestinal organoids in three-dimensional niche in vitro / 中国组织工程研究

BACKGROUND: Induced pluripotent stem cells (iPSCs) are a special type of cells with self-renewal and multi-differentiation potential, which can differentiate into intestinal organoids under certain conditions. OBJECTIVE: To explore whether iPSCs can differentiate into intestinal organoids under specific conditions in vitro.METHODS: iPSCs from B6J mice were recovered and cultured for 3 days until clone units covered about 80％ of the culture dish, and then the cells were cultured in the medium containing Activin A for 3 days until the deterministic endoderm formed. Further, the culture medium was replaced by the medium with fibroblast growth factor 4 and Wnt3A for 4 days to differentiate into the spheroids with CDX2+. After that, spheroids were collected and mixed with Matrigel,and then the mixture was dropped into the 4-well plate and cultured with Rspondin1, Noggin, epidermal growth factor, B27 and other growth factors to differentiate into intestinal organoids. Cell morphology was observed, FoxA2 and Sox17 expresson in the deterministic endoderm was detected, and CDX2, Sox9, CGA, MMP7 were measured.RESULTS AND CONCLUSION: iPSCs were cultured with Activin A for 3 days with higher cell fusion, initial differentiation and FoxA2/Sox17 expression (P < 0.05) than those of non-induced iPSCs. Spheroids began to appear at the 3rd day after culture with fibroblast growth factor 4 and WNT3A, and formed a lot at the 4th day. And CDX2 expression in spheroids was significantly increased compared with that in the deterministic endoderm (P < 0.05). Organoids gradually formed after 3 days culture, which contained all cell types of intestinal organoids, and expressions of specific markers, Sox9, CGA, MMP7, were significantly higher than those in spheroids (P < 0.05). To conclude, iPSCs can be induced to differentiate into intestinal organoids in three-dimensional niche in vitro.

Value of Sysmex UF-1000i Urinary Sediment Analyzer in Diagnosis of Urinary Tract Infection due to Different Urinary Specimen Collection Methods / 医学研究杂志

Objective To explore the value of Sysmex UF-1000i urinary sediment analyzer in diagnosis of urinary tract infection (UTI) and clarify the influence caused by different specimen on UF-1000i diagnosing UTI.Methods Totally 466 specimens examined bacterial culture and routine urinalysis were collected from patients suspected of UTI during July and August,2015 (samples of group A).148 specimens during late March and early April were gathered to implement a urine culture and then the rest of urine were detected byUF -1000i urinary sediment analyzer instantly(samples of group B).Bacteria and leukocyte counts were gathered and then receiver operating characteristic (ROC) carve was drawn regarding thegold standard as bacterial culture by SPSS18.0.Next,the threshold values of bacteria and leukocyte counts for diagnosis of UTI were found out.Meanwhile,itssensitivity,specificity,positive/negative predictive value,false positive/false negative value,and diagnostic accuracy were calculated.Results The cut off values to samples of group A were101.7 bacteria/μl and 18.8WBC/μl respectively and to samples of group B were 98.7 bacteria/μl and 11.1WBC/μl respectively.The area of Bacteria and leukocyte counts under ROC carve was 0.604 and 0.661 to samples of group A and 0.941 and 0.848 to samples Of group B.To samples of group B,combined Bacteria and leukocyte counts for UTI,the optimum sensitivity was 82.4％,specificity was 92.1％,positive predictive value was 77.8％,negative predictive value was 93.8％,false positive rate was 7.9％,false negative rate was 17.6％,and accuracy was 89.9％.Bacterial culture was reduced by 70.9％.Conclusion UF-1000i urine sediment analyzers have the value of early screening value and help to diagnose UTI.Urine that was sterilely collected and examined within two hours can make the value of UF-1000i maximized.

Effects of Laparoscopy Assisted Complete Mesocolon Resection on the Stress Response and Immune Function of Patients with Colon Cancer / 现代生物医学进展

Objective:To investigate the effects of laparoscopy assisted complete mesocolon resection on the stress response and immune function of patients with colon cancer.Methods:From January 2013 to January 2016,130 patients with colon cancer in our hospital were selected and randomly divided into the observation group and the control group with 65 cases in each group according to the random lottery envelopes.The observation group was treated with laparoscopy assisted complete mesocolon resection,the control group was treated with open operation.The changes of semm IL-6,TNF-α and amours of CD4+ cells,blood loss,bowel function restoration time,hospital stays and complication rate of two groups were compared before and after treatment.Results:Both groups of patients were successfully completed the surgery and there was no serious complications occurred in both groups.The amount of bleeding,postoperative intestinal recovery time and postoperative hospital stay in the observation group were significantly less than those of the control group (P＜0.05).The chylous fistula,wound infection,pulmonary infection and complication rate on the 14th day postoperation in the observation group was 1.5％,which was 12.3％ in the control group and significantly higher than that of the control group (P＜0.05).The contents ofCD4+ cells in the observation group and the control group were 36.34± 7.83％ and 33.66± 9.82％,respectively,which were significantly higher than those before treatment(30.33± 6.49％ and 30.49± 5.33％)(P＜0.05),and the contents of CD4+ cells of observation group was significantly higher than that of the control group after treatment (P＜0.05).The serum levels ofIL-6 and TNF-α in both groups were significantly lower than those before treatment on the 14th day postoperation (P＜0.05),and the serum IL-6 and TNF-α levels of observation group were significantly lower than those in the control group on the 14th day postoperation (P＜0.05).Conclusion:Laparoscopy assisted complete mesocolon resection could promote the immune function and inhibit the stress response,which further promoted the rehabilitation and induced the complications in the treatment of patients with colon cancer.

Cytopathic effect of rotavirus NSP4 on rat neurons and its influence on intracellular calcium homeostasis / 中华微生物学和免疫学杂志

Objective To investigate the cytopathic effect of amino acid residues 86 to 175 of rotavirus nonstructural protein 4 (NSP486-175) on rat neurons and to analyze the underlying mechanism.MethodsPrimary cultured rat neurons were treated with NSP486-175 and the morphological changes induced in rat neurons were observed.Lactate dehydrogenase (LDH) activity in the culture supernatant of NSP486-175 treated-neurons was measured.Laser scanning confocal microscope was used to detect the concentration of intracellular Ca2+ labeled with Fluo-3 AM.Results Exogenous addition of NSP486-175 induced obvious cytopathic effect on rat neurons.The LDH activity in the culture supernatant of treated-neurons was stronger than that of the control group.The intensity and the distribution of fluorescence in neurons were altered after stimulation with NSP486-175.Conclusion NSP486-175 can induce the damage of rat neurons, which may be related to its role in increasing the concentration of intracellular Ca2+.This study may provide certain theoretical basis for understanding extra-intestinal spread and pathogenesis of rotavirus infection.

Treatment and prognosis of 76 patients with gallbladder cancer: a single-center retrospective study / 中华肝胆外科杂志

Objective To investigate effective treatment modalities and the related factors influencing prognosis of patients with gallbladder cancer.Methods The clinical data of 76 gallbladder carcinoma patients admitted to the Department of General Surgery,PLA Nanjing General Hospital from January 2005 to October 2015 were analyzed retrospectively.Follow-up was carried out via telephone or outpatient service until January 2016.Cox regression and Kaplan-Meier models were performed for survival analysis.Results 69 patients were treated with surgery and/or postoperative adjuvant chemotherapy.The remaining 7 patients with liver or distant metastases who did not undergo surgery received chemotherapy.24 patients died from cancer relapse,37 patients died from disease progression after giving up treatment,and 7 patients were lost to follow-up.The remaining 8 patients were still alive at the time of follow-up.The depth of cancer invasion (HR =2.736),the type surgical procedure (HR =2.207),and adjuvant chemotherapy (HR =0.603) were significant impact factors of survival for GBC patients.Adjuvant chemotherapy was a protective factor.The average survival in the chemotherapy-naive group was (10.6 ± 1.9) months,the single chemotherapy group (18.5 ± 2.8) months,and the combined chemotherapy group (26.9 ± 6.4) months.There were no significant differences among these groups.Conclusions The depth of cancer invasion,types of surgical procedure particularly radical cholecystectomy,and adjuvant chemotherapy were significant factors of survival in patients with GBC.Radical cholecystectomy combined with arterial and intravenous chemotherapy using gemcitabine and oxaliplatin showed benefits in survival in GBC patients.

"Physical dose estimation for the patient in early stage of ""5.7"" 192Ir source accident in Nanjing" / 中华放射医学与防护杂志

Objective To make a rapid dose estimation for a patient exposed locally to high dose radiation exposure in early stage of 5.7 192Ir source accident in Nanjing.Methods Based on source parameters,exposure pattern,and time duration,the doses were estimated using MCNP simulations with the aid of the proper East Asia adult male voxel phantom and main physiological parameters of the exposed patient.Result Absorbed doses to 16 organs or tissues were estimated to be in the range 0.03 to 9.16 Gy.Also,the iso-dose curves for the skin of legs showed clearly the dose difference between right and left legs.Absorbed doses to patient's testicles and prostrate,about 9.16 Gy,were higher than those to other organs or tissues.The skin of both legs was exposed locally to high dose radiation exposure,for which the estimated doses were in agreement with the results obtained from infrared thermal imaging.Conclusions Monte Carlo simulation technique and common software can be used for dose estimation in early stage of radiation accidents effectively with integrating proper models.

Effect of terlipressin on hepatic and renal function in cirrhotic patients undergoing hepatectomy / 国际外科学杂志

Objective To investigate the effect of terlipressin on hepatic and renal function in cirrhotic patients undergoing hepatectomy.Methods Aanlyze the clinical data of 57 patients following irregular hepatectomy for hepatocellular carcinoma with cirrhosis,according to whether use terlipressin or not after operation,which were divided into terlipressin group (A group,n =27) and control group (B group,n =30).Liver function parameters (ALT,AST,TB),ascites,urine volume and renal function parameters (Cr,BUN) preoperatively and on postoperative day(POD) 1,3,5 and 7 were compared between the two gruops.Results Compared with those of POD 1,the levels of ALT,AST and ascites on POD 3,5,7 were significantly lower in two groups (P ＜ 0.05),urine volume was significantly increased (P ＜ 0.05),Cr of POD 7 was significantly lower (P ＜0.05),but it is more remarkable in group A than group B.The levels of ALT in terlipressin group on POD 5,7 were (144.9 ±76.3) U/L,(100.5 ±61.5) U/L,which were lower than those of (267.2±91.2) U/L,(199.3 ±70.5) U/L in control group.On POD 3,5,7,the levels of AST,BUN,Cr and peritoneal fluid in terlipressin group,which were respectively(211.1 ±99.8) U/L,(80.4 ±54.6) U/L,(50.6 ±46.5) U/L,(6.6 ± 1.9) mmol/L,(6.5 ± 1.7) mmol/L,(6.3 ± 2.1) mmol/L,(74.3 ± 10.9) μmol/L,(71.5 ± 8.9) μmol/L,(58.7 ±4.1) μmol/L,(247.6±60.3) ml,(58.8±54.3) ml,(40.2±31.8) ml,were significantly lower than those in control group which were (298.7 ±131.2) U/L,(201.1 ±93.4) U/L,(114.7 ±70.3) U/L,(7.3 ± 1.9) mmol/L,(7.2±1.8) mmol/L,(7.1±1.7) mmol/L,(79.5±15.1) μmol/L,(76.9±16.2) μmol/L,(69.4±11.4) μmol/L,(275.2±88.1) ml,(191.7±71.6) ml,(93.2±50.2) ml.while urine volume of (2232.3±409.8) ml,(2270.5 ±395.8) ml,(2179.0±301.4) ml was much more than that of (1921 ± 510.4) ml,(2019.1 ±411.2) ml,(1978.7±323.7) ml in the control group,the differences in the two groups were statistically significant (P ＜ 0.05).There were 11 (36.7％) patients with hepatic and renal dysfunction and hepatorenal syndrome after operation in group B,while only 2 (7.4％) patients in group A.Conclusions The use of terlipressin after partial liver resection has a protective effect on hepatic and renal function in patients with cirrhosis,and can reduce postoperative ascites and prevent hepatorenal syndrome.

Comparisons of DNA damages in physics process caused by protons with different energies / 中华放射医学与防护杂志

Objective To compare the difference among DNA damages in physics processes caused by protons with different energies around Bragg peak.Methods By constructing the nucleus and DNA model using Geant4 and simulating the transportation processes of protons with different energies around Bragg peak in nucleus model based on Geant4-DNA,the information of interaction points were recorded.16％ of them were picked randomly to use as the points at which protons interact with DNA.After finding out the points which broke the DNA and writing their information to new files,these new files were treated with density-based spatial clustering of applications with noise (DBSCAN),so as to analyze and calculate the difference of DNA damages caused by protons with different energies.Results With the protons with energies from 20.0 to 0.6 MeV,the numbers of damage points and damage clusters increased from 49.86 to 549.88 and from 2.92 to 82.46,respectively;the numbers of clusters with different sizes had a remarkable increase,and the number of clusters with size not less than 5 increased by 400 times.The average cluster size increased slightly.Simple single strand breaks (SSSBs),complex single strand breaks (CSSBs),simple double strand breaks (SDSBs) and complex double strand breaks (CDSBs) of DNA increased by about 8,26,24 and 64 times,respectively.The proportion of single strand breaks (SSBs) decreased from 96.69％ to 89.37％,and the double strand breaks (DSBs) increased from 3.31％ to 10.63％.Conclusions The lower proton energy would result in the more complex damage to DNA and the harder repair of DNA damage.