Early colorectal cancer detection: a serum analysis platform combining SERS and machine learning.

Colorectal cancer (CRC) is one of the deadliest malignancies globally, with high incidence and mortality rates. Early detection is crucial for improving treatment success rates and patient survival. However, due to the difficulty in detecting early symptoms, many cases are diagnosed at advanced stages, necessitating more sensitive and accurate detection methods. This study proposes a novel approach combining the Principal Component Analysis (PCA)-Dynamic Weighted Nearest Neighbor (DWNN) model with Surface-Enhanced Raman Scattering (SERS) technology to detect the serum of CRC mice at different stages. Establishing the CRC mice model, serum samples were collected for further analysis. An Au Nanocluster (AuNC) substrate was synthesized to ensure optimal SERS enhancement. The PCA-DWNN recognition model was constructed to classify the SERS spectra of CRC at different stages. The synthesized AuNC substrate has high sensitivity, good reproducibility, uniformity, and stability, making it a high-performance nanomaterial. The PCA-DWNN model has significant advantages in identifying high-dimensional and complex SERS spectra, offering excellent classification accuracy and robustness, with an accuracy rate of 97.5%. By analyzing the PCA loading plot, it was observed that as CRC progressed, the content and structure of proteins, lipids, amino acids, and carbohydrates in the serum changed, reflected in different characteristic peaks in the SERS spectra. This study suggests that SERS combined with PCA-DWNN has potential in the early detection of CRC, possibly providing a novel approach for clinical diagnostics.

Association between serum triglyceride level and severity of acute biliary pancreatitis.

BACKGROUND: Acute biliary pancreatitis (ABP) is the most common type of acute pancreatitis. However, the effect of serum triglyceride (TG) levels on the severity of ABP remains unclear. The aim of this study was to assess the correlation between serum TG levels and the severity of ABP. METHODS: Data from 526 ABP patients was analyzed in this study. The patients were divided into normal and elevated groups according to the TG level measured within 24 h after admission, and the elevated group was further divided into mild, moderate, and severe elevated groups. The demographic data and clinical outcomes of each group were compared. RESULTS: Of the 526 ABP patients, 394 were in the normal TG group and 132 were in the elevated TG group (36 mild, 57 moderate, and 39 severe). The elevated group was younger (51.5 ± 12.9 vs. 58.9 ± 13.9), predominantly male (66.7% vs. 45.2%), had more history of diabetes (22.7% vs. 12.4%) and hyperlipidemia (19.7% vs. 0.8%), and developed systemic inflammatory response syndrome (SIRS) (25.8% vs. 15.5%), persistent organ failure (POF) (11.4% vs. 2.8%), and local complications (62.9% vs. 42.1%) more frequently compared to the normal group (P < 0.05). The incidence of SIRS, POF, acute peripancreatic fluid collection (APFC), and acute necrotic collection (ANC) increased with increasing TG levels (Ptrend < 0.05). In multivariate analysis, TG was independently associated with POF, APFC, and ANC in increments of 100 mg/dl (P < 0.05), and there was a linear relationship between TG levels and POF, APFC, and ANC (non-linear P > 0.05, P overall <0.05). In addition, nonalcoholic fatty liver disease is not a risk factor for POF, ANC, and APFC in ABP patients. CONCLUSIONS: Elevated serum TG levels were independently associated with more severe ABP. The incidence of POF, APFC, and ANC in ABP patients increased with the increase of TG levels, with a linear relationship.

Traumatic biliary fistula with bile duct disconnection.

A 43-year-old man was admitted to our department for accident. Computed tomography (CT) scan revealed low-density shadows in the hepatogastric and peripancreatic space. The patient still has abdominal pain after the performation of CT-guided abdominal puncture and drainage. Magnetic Resonance Cholangiopancreatography (MRCP) showed that the middle and lower segments of common bile duct were stricture and truncature. Endoscopic bile duct catheterization and drainage showed that the lower segment was dislocated and disconnected. Then the doctor used the balloon and contrast agent to pressurize, and at the same time, the distal common bile duct was pulled by the mirror, trying to straighten the common bile duct, changing its axial direction, and try to bridge it repeatedly by using the guide wire of misgurnus anguillicaudatus. The patient significantly improved over the next 3 days. After 11 months of follow-up, the patient came to our hospital to remove the biliary stent,and has no abdominal pain anymore.

Relation between triglycerides and the severity of acute pancreatitis combined with nonalcoholic fatty liver disease: a retrospective study.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) can exacerbate the severity of acute pancreatitis (AP), and this severity is worsened with increased severity of NAFLD. This study aimed to investigate the relation between serum triglyceride (TG) and the severity of AP with NAFLD by collecting clinical data from AP patients with NAFLD. METHODS: AP patients with NAFLD were divided into 2 groups according to TG levels: hypertriglyceridemia (HTG) group and non-hypertriglyceridemia (NHTG) group. RESULTS: In total, 598 AP patients with NAFLD were enrolled in this study, including 433 in the HTG group and 165 in the NHTG group. Compared with the NHTG group, AP patients in the HTG group were more serious (P < 0.05). The incidence of persistent organ failure (POF), especially persistent respiratory failure, and the ratio of acute peripancreatic fluid collection (APFC) were higher in the HTG group (P < 0.05). Higher TG levels were associated with a higher incidence of APFC (P < 0.05). Logistic regression analysis showed that the risk of APFC was significantly higher in moderate and severe NAFLD than in mild NAFLD. CONCLUSION: HTG may aggravate the severity and local complications of AP combined with NAFLD.

Nitrogen isotopes suggest agricultural and non-agricultural sources contribute equally to NH3 and NH4+ in urban Beijing during December 2018.

Agricultural and non-agricultural sources emission contribute to atmospheric ammonia (NH3) and particulate ammonium (NH4+). However, our understanding on the sources of NH3 and NH4+ in PM2.5 (particles smaller than 2.5 µm) during the winter period in the urban atmosphere is limited. Here, we measured the concentrations and stable nitrogen isotopic composition (δ15N) of NH3 and NH4+ in parallel during December 2018 in urban Beijing to assess the non-agricultural and agricultural sources contributions to NH3 and NH4+ in ambient air based on the Chemical Transport Model (CTM), a Bayesian isotope mixing model (SIMMR), and the δ15N signatures that we developed. Our study found weekly NH4+ and NH3 concentrations were on average 2.5 ± 1.4 µg m-3 and 3.8 ± 1.7 µg m-3, respectively during December 2018. Weekly concentration weighted δ15N(NH4+) values ranged from 4.5‰ to 13.7‰ with an average value of 8.2 ± 3.9‰ during December 2018. After accounting for nitrogen isotopic fractionation from NH3 gas to NH4+ conversion, initial δ15N(NH3) values ranged from -22.5‰ to -12.8‰ with an average value of -17.4 ± 3.5‰. Further, weekly measured δ15N(NH3) values ranged from -22.2‰ to -10.2‰ (after correction) with an average value of -15.6 ± 5.3‰ during December 2018. Results from two different isotope-based method showed non-agricultural sources contributed 31.2%-63.1%, with an average value of 47.5 ± 14.6%, to NH4+ and 32.3%-71.2%, with an average of 53.4 ± 16.1%, to ambient NH3 during December 2018 in Beijing. Results from CMAQ-ISAM suggest non-agricultural sources contributed on average 66.2 ± 1.9% to ambient NH4+ and 66.4 ± 1.9% to ambient NH3 during December 2018. Results from this study suggest that agricultural and non-agricultural sources nearly equally contributed to NH3 and NH4+ in urban Beijing during December 2018 with an uncertainty range of 13%-19% between isotope-based methods and CTM method.

δ15N-stable isotope analysis of NH x : An overview on analytical measurements, source sampling and its source apportionment.

Agricultural sources and non-agricultural emissions contribute to gaseous ammonia (NH3) that plays a vital role in severe haze formation. Qualitative and quantitative contributions of these sources to ambient PM2.5 (particulate matter with an aerodynamic equivalent diameter below 2.5 µm) concentrations remains uncertain. Stable nitrogen isotopic composition (δ15N) of NH3 and NH4 + (δ15N(NH3) and δ15N(NH4 +), respectively) can yield valuable information about its sources and associated processes. This review provides an overview of the recent progress in analytical techniques for δ15N(NH3) and δ15N(NH4 +) measurement, sampling of atmospheric NH3 and NH4 + in the ambient air and their sources signature (e.g., agricultural vs. fossil fuel), and isotope-based source apportionment of NH3 in urban atmosphere. This study highlights that collecting sample that are fully representative of emission sources remains a challenge in fingerprinting δ15N(NH3) values of NH3 emission sources. Furthermore, isotopic fractionation during NH3 gas-to-particle conversion under varying ambient field conditions (e.g., relative humidity, particle pH, temperature) remains unclear, which indicates more field and laboratory studies to validate theoretically predicted isotopic fractionation are required. Thus, this study concludes that lack of refined δ15N(NH3) fingerprints and full understanding of isotopic fractionation during aerosol formation in a laboratory and field conditions is a limitation for isotope-based source apportionment of NH3. More experimental work (in chamber studies) and theoretical estimations in combinations of field verification are necessary in characterizing isotopic fractionation under various environmental and atmospheric neutralization conditions, which would help to better interpret isotopic data and our understanding on NH x (NH3 + NH4 +) dynamics in the atmosphere. ELECTRONIC SUPPLEMENTARY MATERIAL: Supplementary material is available in the online version of this article at 10.1007/s11783-021-1414-6 and is accessible for authorized users. Supplementary material includes supplementary tables on summary of recent isotope-based source apportionment studies on ambient NH3 derived from δ15N(NH3) values (Table A1); and summary of recent isotope-based source apportionment studies on particulate NH4 + derived from δ15N(NH4 +) values (Table A2).

Polar organic aerosol tracers in two areas in Beijing-Tianjin-Hebei region: Concentration comparison before and in the sept. Third Parade and sources.

A total of 106 24-h PM2.5 aerosol samples were collected in an urban area (Shijiazhuang, SJZ) and a suburban area (Liulihe, LLH, Fangshan County, Beijing) in the Beijing-Tianjin-Hebei (BTH) region in 2 periods: the first is from 10 July to 10 August, which is before Sept. Third Parade (Period I); the second is from 20 Aug. to 6 Sept. 2015, which is during Sept. Third Parade (Period II). Polar organic tracers, including isoprene, α-pinene, ß-caryophyllene and toluene oxidation products, as well as sugars and carboxylic acids were measured. In Period II, rigorous emission-reduction measures were taken in the BTH region. With the anthropogenic emission being cut down significantly, the average concentrations of isoprene, α-pinene, ß-caryophyllene and toluene oxidation products and all carboxylic acids (except tetradecanoic, palmitic, and stearic acids), were lower in Period II than those in Period I in LLH, indicating that the SOA tracers were decreased with precursor emission volumes and yields in the atmosphere. Moreover, sugar compounds were shown with comparable levels during the two periods in LLH, suggesting that no measures were taken to reduce the intensities of the biogenic sources. On the contrary, tetradecanoic, palmitic, and stearic acids were shown with obviously higher concentrations in Period II than those in Period I, demonstrating that cooking fumes increased during Sept. Third Parade period. The positive matrix factorization (PMF) model combining with tracer-based method was applied to explore the sources of secondary organic carbon (SOC). It reveals that the sources of SOC include isoprene, α-pinene, ß-caryophyllene and toluene oxidation products, fossil fuel combustion, cooking fumes and regionally transferred aged aerosols. These sources accounted for 11.3%, 9.0%, 15.5%, 10.9%, 29.2%, 2.9%, 21.1% of SOC for SJZ, and 12.7%, 11.2%, 9.7%, 14.4%, 25.3%, 0%, 26.7% of SOC for LLH, during the whole sampling periods respectively.

Sources of gaseous NH3 in urban Beijing from parallel sampling of NH3 and NH4+, their nitrogen isotope measurement and modeling.

Atmospheric gaseous ammonia (NH3) is the most abundant alkaline gas in the atmosphere while aerosol ammonium (NH4+) constitutes a majority of the inorganic cation concentration in total PM2.5 mass and plays a vital role in severe haze formation. This study tried to shed some light on sources of gaseous NH3 through integrating the parallel measurements of δ15N values in NH4+ and ambient NH3, NH3 source signature measurement, IsoSource model, and chemistry and transport model (CTM). As a result, predicted initial δ15N (NH3) values ranging from -42.0‰ to -4.9‰ were derived from daily δ15N(NH4+) values of fine particulate NH4+, and δ15N(NH3) values ranging from -26.8‰ to -17.2‰ were obtained from weekday/weekend δ15N(NH3) values, respectively. During summer, non-agricultural sources (e.g. fossil fuel sources, urban waste) contributed 63% to ambient NH3 in urban Beijing, derived from δ15N(NH3) values whereas 64% to ambient NH3, derived from δ15N(NH4+) values. These results suggested that non-agricultural sources were main contributors to gaseous NH3 even during summer and agricultural sources were not likely the main source of gaseous NH3 in urban Beijing. To further reduce the uncertainty of isotope-based source apportionment results, more laboratory and field studies are necessary to refine the δ15N(NH3) source profile of NH3 using validated collection technique as overlapping exist between δ15N(NH3) values in agricultural sources such as livestock breeding waste (-42.5‰ to -29.1‰) and fertilizer application (-51.5‰ to -35.0‰).

Chemical characteristics and sources of water-soluble organic aerosol in southwest suburb of Beijing.

PM2.5 filter sampling and components measurement were conducted in autumn and winter from 2014 to 2015 at a suburban site (referred herein as "LLH site") located in the southwest of Beijing. The offline aerosol mass spectrometry (offline-AMS) analysis and positive matrix factorization (PMF) were applied for measurement and source apportionment of water-soluble organic aerosol (WSOA). Organic aerosol (OA) always dominated PM2.5 during the sampling period, especially in winter. WSOA pollution was serious during the polluted period both in autumn (31.1 µg/m3) and winter (31.9 µg/m3), while WSOA accounted for 54.4% of OA during the polluted period in autumn, much more than that (21.3%) in winter. The oxidation degree of WSOA at LLH site was at a high level (oxygen-to-carbon ratio, O/C=0.91) and secondary organic aerosol (SOA) contributed more mass ratio of WSOA than primary organic aerosol (POA) during the whole observation period. In winter, coal combustion OA (CCOA) was a stable source of OA and on average accounted for 25.1% of WSOA. In autumn, biomass burning OA (BBOA) from household combustion contributed 38.3% of WSOA during polluted period. In addition to oxygenated OA (OOA), aqueous-oxygenated OA (aq-OOA) was identified as an important factor of SOA. During heavy pollution period, the mass proportion of aq-OOA to WSOA increased significantly, implying the significant SOA formation through aqueous-phase process. The result of this study highlights the concentration on controlling the residential coal and biomass burning, as well as the research needs on aqueous chemistry in OA formation.

Transcutaneous Electrical Acustimulation Improves Gastrointestinal Disturbances Induced by Transcatheter Arterial Chemoembolization in Patients With Liver Cancers.

BACKGROUND: Gastrointestinal (GI) disturbances occur in patients who receive chemotherapy via transcatheter arterial chemoembolization (TACE) and could last for an extended period of time in some cases. Antiemetic drugs have a potential risk of developing hepatic failure and are ineffective for delayed nausea and emesis. Transcutaneous electrical acustimulation (TEA) has recently been reported to exert antiemetic and prokinetic effects, but it is unknown whether it has an ameliorating effect on TACE-induced GI disturbances. AIM: This study was designed to evaluate effects and mechanisms of noninvasive TEA on GI symptoms in patients treated with TACE. MATERIALS AND METHODS: Seventy-four patients with liver cancers (eighteen female; age 63.4 ± 1.1 years) scheduled for TACE were randomized to TEA (n = 37) or sham-TEA (n = 37). TEA was performed via acupoints, ST36 and PC6 using parameters previously optimized for GI motility (1 h, bid) from the postoperative day 0 (POD0) to POD2. Sham-TEA was performed using the same parameters via non-acupoints. Symptom questionnaires were completed daily. The electrogastrogram (EGG) and electrocardiogram (ECG) were recorded in the fasting state for 30 mins to assess gastric slow waves and autonomic functions, respectively, before and after the 3-day treatment. RESULTS: 1) In the acute phase (<24 h), TEA showed no effects on any of GI symptoms, compared with sham-TEA. 2) In the delayed phase (>24 h), TEA, compared with sham-TEA, decreased the percentage of patients who experienced nausea on POD3 (0% vs. 13.5%, p = 0.021), the nausea score on POD3 (p = 0.022), the anorexia score on POD2 (p = 0.040) and POD3 (p = 0.004), and the bloating score (POD1-3: p < 0.01). 3) In comparison with sham-TEA, TEA increased the number of spontaneous bowel movements (p = 0.001) and the Bristol score of the first stool (p = 0.014) and decreased the number of patients with the use of laxatives (p = 0.022). 4) Physiologically, the 3-day TEA but not sham-TEA increased the percentage of normal gastric slow waves (p < 0.001) and vagal activity (p = 0.006). The vagal activity was negatively correlated with the anorexia score (r = -0.267, p = 0.026). It was found that the sympathovagal ratio and tumor size>5 cm were independent risk factors predicting the occurrence of nausea in patients after TACE. CONCLUSION: TEA improves major TACE-induced GI disturbances in the delayed phase, including nausea, bloating, impaired gastric pace-making activity, and constipation in patients with liver cancers via the autonomic pathway.

Nitrate dominates the chemical composition of PM2.5 during haze event in Beijing, China.

Water-soluble inorganic ions (WSI), a major component of PM2.5, often increased rapidly during the haze event in Beijing. Sulfate (SO42-), Nitrate (NO3-), and Ammonium (NH4+) are three main components of WSI. Since year 2015, sulfate concentrations in PM2.5 has gradually decreased owing to the effective control of SO2 emissions. However, the contribution of nitrate to PM2.5 has significantly increased during haze events in Beijing at the same time. In this study, a highly time-resolved online analyzer (Monitor for Aerosols and Gases, MARGA) was employed to measure the WSI in PM2.5 in Beijing from 5 February to 15 November 2017. Three typical haze events during this sampling period were investigated. During heavy pollution episodes in winter, nitrate concentrations increased from 7.5 µg/m3 to 45.6 µg/m3 (45.0% of WSI), while sulfate increased from 4.2 µg/m3 to 20.1 µg/m3 (19.8% of WSI). This indicated that nitrate is more important than sulfate as a driver for the growth of PM2.5 during the period of heavy air pollution in winter. Nitrate also dominates the increase of WSI in the pollution episodes in autumn, with an average concentration of 52.5 µg/m3, and contributed up to 67% of WSI. The average concentration ratio of NH4+ to SO42- was higher in autumn (1.02) than that in summer (0.74) and close to that in winter (1.00). This is mainly because the emission control of coal combustion in Beijing and surrounding areas results in an NH3-rich and SO2-lean atmosphere, which promoted the formation of ammonium nitrate. Our study indicates that nitrate has become the most important component of WSI in PM2.5 and is driving the rapid growth of PM2.5 concentrations during heavy pollution episodes in Beijing. Therefore, more efforts shall be made to reduce the nitrogen oxide and ammonia emissions in Beijing and surrounding areas.

[Source Apportionment of PM2.5 in Suburban Area of Beijing-Tianjin-Hebei Region in Autumn and Winter].

To identify the main sources of PM2.5 in Beijing-Tianjin-Hebei (BTH) region, PM2.5 samples were collected at four suburban sites in BTH region during autumn and winter in 2014-2015. Source apportionment of PM2.5 was conducted using the chemical mass balance model (CMB). It shows that the main sources of PM2.5 in autumn and winter were secondary aerosols (36%-58%), traffic (8%-26%), residential coal combustion (8%-16%), and biomass burning (5%-16%). Secondary nitrate was the most important source of PM2.5 at most sites during autumn and winter (11%-27%). The source apportionment at different pollution levels indicates that the coherence of the increasing trend of different sources among the four sites were much more obvious in autumn than in winter. Also, the increasing contribution of secondary sources (47.2-115.7 µg·m-3) was much higher than that of primary sources (29.5-43.4 µg·m-3) in autumn, but such trend was not significant in winter. The total contribution of coal combustion at suburban sites was quite similar to that in urban sites, but in suburban areas residential coal combustion dominates the contribution from coal combustion. Thus, it is very necessary for suburban areas of the BTH region to control emissions from residential coal combustion.

Molecular Analysis-Based Genetic Characterization of a Cohort of Patients with Duchenne and Becker Muscular Dystrophy in Eastern China.

BACKGROUND: Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are common X-linked recessive neuromuscular disorders caused by mutations in dystrophin gene. Multiplex polymerase chain reaction (multiplex PCR) and multiplex ligation-dependent probe amplification (MLPA) are the most common methods for detecting dystrophin gene mutations. This study aimed to contrast the two methods and discern the genetic characterization of patients with DMD/BMD in Eastern China. METHODS: We collected 121 probands, 64 mothers of probands, and 15 fetuses in our study. The dystrophin gene was detected by multiplex PCR primarily in 28 probands, and MLPA was used in multiplex PCR-negative cases subsequently. The dystrophin gene of the remaining 93 probands and 62 female potential carriers was tested by MLPA directly. In fetuses, multiplex PCR and MLPA were performed on 4 fetuses and 10 fetuses, respectively. In addition, sequencing was also performed in 4 probands with negative MLPA. RESULTS: We found that 61.98% of the subjects had genetic mutations including deletions (50.41%) and duplications (11.57%). There were 43.75% of mothers as carriers of the mutation. In 15 fetuses, 2 out of 7 male fetuses were found to be unhealthy and 2 out of 8 female fetuses were found to be carriers. Exons 3-26 and 45-52 have the maximum frequency in mutation regions. In the frequency of exons individually, exon 47 and exon 50 were the most common in deleted regions and exons 5, 6, and 7 were found most frequently in duplicated regions. CONCLUSIONS: MLPA has better productivity and sensitivity than multiplex PCR. Prenatal diagnosis should be applied in DMD high-risk fetuses to reduce the disease incidence. Furthermore, it is the responsibility of physicians to inform female carriers the importance of prenatal diagnosis.

Characteristics and sources of aerosol pollution at a polluted rural site southwest in Beijing, China.

Annual average PM2.5 concentration in south Beijing was 30% and 40% higher than the whole Beijing city in 2015 and 2016, respectively. Few studies have been conducted to investigate what leads to the characteristics and sources of heavy pollution in the south rural area of Beijing. This study conducted an observation with Aerosol Chemical Speciation Monitor (ACSM) at a southwest rural site (Liulihe) in Beijing during 2014-2016, to investigate the seasonal aerosol characteristics and their sources. Positive matrix factorization (PMF) algorithm was used to distinguish different components of organic aerosol measured by ACSM. Biomass burning is an important pollution source, mainly due to the open burning after harvest season in autumn, regional transport in spring, and local residential biofuel use in winter. Coal consumption is the largest primary organic aerosol source in winter. Heavy duty diesel trucks contributed significantly to organic aerosol at night-time in the rural area. Results of this study show residential solid fuel burning is the most important source of aerosol pollution in the rural area of Beijing and the results focusing on urban Beijing might have underestimate the contribution from residential emissions in the Beijing-Tianjin-Hebei region.

Galectin-1-driven upregulation of SDF-1 in pancreatic stellate cells promotes pancreatic cancer metastasis.

Galectin-1, mainly expressed in activated pancreatic stellate cells (PSCs), is involved in many important cancer-related processes. However, very little is known how Galectin-1 modulates PSCs and subsequently impacts pancreatic cancer cells (PCCs). Our chemokine antibody array and in vitro studies demonstrates that Galectin-1 induces secretion of stromal cell-derived factor-1(SDF-1) in PSCs by activating NF-κB signaling. The secreted SDF-1 increases migration and invasion of PCCs. Knockdown of Galectin-1 and inhibitor-mediated blockade of SDF-1 as well as its ligand CXCR4 and NF-κB verifies the findings. In vivo experiment by knockdown of Galectin-1 in PSCs further demonstrates the conclusion. Collectively, the present studies demonstrate that Galectin-1-driven production of SDF-1 in PSCs through activation of NF-κB promotes metastasis in PDAC, offering a potential target in the treatment of pancreatic cancer.

Modified methods for isolation of pancreatic stellate cells from human and rodent pancreas.

Primary cultures of pancreatic stellate cells (PSCs) remain an important basis for in vitro study. However, effective methods for isolating abundant PSCs are currently lacking. We report on a novel approach to isolating PSCs from normal rat pancreases and human pancreatic ductal adenocarcinoma (PDAC) tissue. After anaesthesia and laparotomy of the rat, a blunt cannula was inserted into the pancreatic duct through the anti-mesentery side of the duodenum, and the pancreas was slowly infused with an enzyme solution until all lobules were fully dispersed. The pancreas was then pre-incubated, finely minced and incubated to procure a cell suspension. PSCs were obtained after the cell suspension was filtered, washed and subject to gradient centrifugation with Nycodenz solution. Fresh human PDAC tissue was finely minced into 1×1×1 mm3 cubes with sharp blades. Tissue blocks were placed at the bottom of a culture plate with fresh plasma (EDTA-anti-coagulated plasma from the same patient, mixed with CaCl2) sprinkled around the sample. After culture for 5-10 days under appropriate conditions, activated PSCs were harvested. An intraductal perfusion of an enzyme solution simplified the procedure of isolation of rat PSCs, as compared with the multiple injections technique, and a modified outgrowth method significantly shortened the outgrowth time of the activated cells. Our modification in PSC isolation methods significantly increased the isolation efficiency and shortened the culture period, thus facilitating future PSC-related research.

Effect of antecolic or retrocolic route of gastroenteric anastomosis on delayed gastric emptying after pancreaticoduodenectomy: A meta-analysis of randomized controlled trials.

BACKGROUND: Delayed gastric emptying (DGE) is one of the most troublesome complications after classical pancreaticoduodenectomy (PD) or pylorus-preserving PD. Whether the route of gastroenteric reconstruction has any influence on DGE remains controversial. The aim of this study was to investigate the influence of different types of gastroenteric anastomosis on DGE after PD/PPPD. METHODS: A systematic search of literature databases (Cochrane Library, PubMed, EMBASE, and Web of Science) was performed to identify eligible studies. Cochrane collaboration's tool for assessing risk of bias was utilized to evaluate the quality of included studies. The primary outcome was DGE incidence rate. Further outcomes included mortality, morbidity, and other operation related events. Random-effect or fix-effect models were used as appropriate. RESULTS: Five randomized controlled trials (RCTs) including a total of 530 patients were identified and included in the analysis. Based on these studies, no difference was found in DGE incidence between antecolic and retrocolic groups (relative risk [RR], 0.82; 95% confidence interval [CI], 0.51-1.32; P = 0.41). Mortality, morbidity, and operation related events were not significantly different between groups. CONCLUSIONS: Results of the meta-analysis reveal that DGE occurrence is not affected by route of gastroenteric anastomosis. Anastomosis approach should be chosen according to the surgeons' preference.

Activation of pancreatic stellate cells involves an EMT-like process.

Pancreatic adenocarcinoma (PDAC) and chronic pancreatitis (CP) are characterized by a desmoplastic reaction involving activated pancreatic stellate cells (PSCs). However, the mechanisms of PSC activation remain poorly understood. We examined whether the epithelial-mesenchymal transition (EMT) process might play a role in PSC activation. PSCs were isolated from a rat pancreas and characterized using immunofluorescence and immunocytochemistry. We evaluated changes in cell motility and in the expression levels of a panel of EMT-related genes during the PSC activation process. Activation of PSCs occurred after 48 h of in vitro culture, as indicated by a morphological change to a myofibroblastic shape and a decrease in the number of cytoplasmic lipid droplets. After activation, PSCs showed enhanced cell migration ability compared to quiescent cells. In addition, the expression of epithelial markers (E-cadherin, BMP7 and desmoplakin) decreased, while expression of mesenchymal markers (N-cadherin, vimentin, fibronectin1, collagen1α1 and S100A4) increased in activated PSCs. EMT-related transcription factors (Snail and Slug) were also upregulated after PSC activation. The concurrent increase in cell migration ability and alterations in EMT-related gene expression suggests that the activation of PSCs involves an EMT-like process. The knowledge that PSC activation involves an EMT­like process may help to identify potential new therapeutic targets to alleviate pancreatic fibrosis in diseases like CP and PDAC.

Living T9 glioma cells expressing membrane macrophage colony-stimulating factor produce immediate tumor destruction by polymorphonuclear leukocytes and macrophages via a "paraptosis"-induced pathway that promotes systemic immunity against intracranial T9 gliomas.

Cloned T9-C2 glioma cells transfected with membrane macrophage colony-stimulating factor (mM-CSF) never formed subcutaneous tumors when implanted into Fischer rats, whereas control T9 cells did. The T9-C2 cells were completely killed within 1 day through a mechanism that resembled paraptosis. Vacuolization of the T9-C2 cell's mitochondria and endoplasmic reticulum started within 4 hours after implantation. By 24 hours, the dead tumor cells were swollen and terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL)-positive. Bcl2-transduced T9-C2 cells failed to form tumors in rats. Both T9 and T9-C2 cells produced cytokine-induced neutrophil chemoattractant that recruited the granulocytes into the tumor injection sites, where they interacted with the tumor cells. Freshly isolated macrophages killed the T9-C2 cells in vitro by a mechanism independent of phagocytosis. Nude athymic rats treated with antiasialo GM1 antibody formed T9-C2 tumors, whereas rats treated with a natural killer cell (NK)-specific antibody failed to form tumors. When treated with antipolymorphonuclear leukocyte (anti-PMN) and antimacrophage antibodies, 80% of nude rats formed tumors, whereas only 40% of the rats developed a tumor when a single antibody was used. This suggests that both PMNs and macrophages are involved in the killing of T9-C2 tumor cells. Immunocompetent rats that rejected the living T9-C2 cells were immune to the intracranial rechallenge with T9 cells. No vaccinating effect occurred if the T9-C2 cells were freeze-thawed, x-irradiated, or treated with mitomycin-C prior to injection. Optimal tumor immunization using mM-CSF-transduced T9 cells requires viable tumor cells. In this study optimal tumor immunization occurred when a strong inflammatory response at the injection of the tumor cells was induced.

T9 glioma cells expressing membrane-macrophage colony stimulating factor produce CD4+ T cell-associated protective immunity against T9 intracranial gliomas and systemic immunity against different syngeneic gliomas.

Cloned T9 glioma cells (T9-C2) expressing the membrane form of macrophage colony stimulating factor (mM-CSF) inoculated subcutaneously into rats do not grow and glioma-specific immunity is stimulated. Immunotherapy experiments showed that intracranial T9 tumors present for one to four days could be successfully eradicated by peripheral vaccination with T9-C2 cells. CD4+ and CD8+ T splenocytes from immunized rats, when restimulated in vitro with T9 cells, produced interleukin-2 and -4. Protective immunity against intracranial T9 gliomas could only be adoptively transferred into naive rats by the CD4+ splenocytes obtained from T9-C2 immunized rats. Rats immunized by the T9-C2 tumor cells also resisted two different syngeneic gliomas (RT2 and F98) but allowed a syngeneic NUTU-19 ovarian cancer to grow. Such cross-protective immunity against unrelated gliomas suggests that mM-CSF transfected tumor cells have immunotherapeutic potential for use as an allogeneic tumor vaccine.