Synchronization isolation method for multiple types of cells from mouse liver / 中华肝脏病杂志

Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.

A differential peptidomics analysis of hippocampal tissue in a rat model of premature white matter injury / 中国当代儿科杂志

OBJECTIVE@#To observe differential peptidomics in the hippocampal tissue in a rat model of premature white matter injury, and to investigate the mechanism of premature white matter injury.@*METHODS@#Twenty neonatal Sprague-Dawley rats were randomly and equally divided into a control group and a model group. Rats in the model group underwent permanent ligation of the right common carotid artery 2 days after birth, followed by 2 hours of hypoxia. For rats in the control group, the right common carotid artery was isolated, but without ligation and hypoxia. Brain tissue samples were collected from the two groups, and hippocampal tissue was isolated. Liquid chromatography-tandem mass spectrometry combined with tandem mass spectrometry was used for peptidomic profiling of hippocampal tissue, and the differentially expressed peptides between the two groups were subjected to bioinformatics analysis to assess their possible roles in neural development and function.@*RESULTS@#A total of 4164 peptides were identified and quantified, and 262 of them were differentially expressed (absolute fold change ≥2.5), including 164 upregulated peptides and 98 downregulated peptides. The numbers of differentially expressed peptides of the precursor proteins ELN, PCLO, MYO15a, MAP4, and MAP1b were the most, and may play significant roles in the pathogenesis of premature white matter injury. CDK5 signaling pathway in the hippocampus was activated in the rat model of premature white matter injury.@*CONCLUSIONS@#The differentially expressed peptides related to precursor proteins such as MAP1b may be key bioactive peptides involved in neural development and function in premature white matter injury, and activation of the CDK5 signaling pathway may be associated with premature white matter injury.

The value of CT texture analysis in differentiating autoimmune pancreatitis from pancreatic ductal adenocarcinoma / 实用放射学杂志

Objective To investigate the feasibility of the CT texture analysis (CTTA)in differentiating autoimmune pancreatitis (AIP)from pancreatic ductal adenocarcinoma (PDAC).Methods 25 patients with AIP and 31 patients with PDAC who confirmed by pathological or clinical underwent pretreatment three-phase contrast-enhanced CT were enrolled.Histogram parameters (mean CT values,median CT values,25 th,75 th percentile CT values,skewness,kurtosis,entropy and uniformity)were derived from CT images through texture analysis.The differences of histogram parameters between AIP and PDAC groups were compared.ROC and AUC were used to evaluate the diagnostic efficacy of histogram parameters in differentiating AIP from PDAC.Results The values for mean CT values,median CT values,25 th,75 th percentile CT values and uniformity of AIP were significantly higher than those of PDAC group,while the values for entropy of AIP were significantly lower than those of PDAC group in arterial phase,portal phase,and delay phases (all P＜0.05). There were no significant differences in kurtosis and skewness between AIP and PDAC groups (all P＞0.05).The uniformity in portal phase achieved the optimal diagnostic accuracy in differentiating AIP from PDAC (AUC＝0.973 ),the cutoff value was 0.797,the corresponding sensitivity and specificity were 92% and 9 6.8%,respectively.Conclusion CTTA can be used as a quantitative analysis method for differential diagnosis between AIP and PDAC,providing a reference for clinicians to select therapeutic schedules.

Clinical comparison between a percutaneous hydraulic pressure delivery system and balloon tamp system using high-viscosity cement for the treatment of osteoporotic vertebral compression fractures

OBJECTIVES: Osteoporotic vertebral compression fractures (OVCFs) affect the elderly population, especially postmenopausal women. Percutaneous kyphoplasty is designed to treat painful vertebral compression fractures for which conservative therapy has been unsuccessful. High-viscosity cement can be injected by either a hydraulic pressure delivery system (HPDS) or a balloon tamp system (BTS). Therefore, the purpose of this study was to compare the safety and clinical outcomes of these two systems. METHODS: A random, multicenter, prospective study was performed. Clinical and radiological assessments were carried out, including assessments of general surgery information, visual analog scale, quality of life, cement leakage, and height and angle restoration. RESULTS: Using either the HPDS or BTS to inject high-viscosity cement effectively relieved pain and improved the patients' quality of life immediately, and these effects lasted at least two years. The HPDS using high-viscosity cement reduced cost, surgery time, and radiation exposure and showed similar clinical results to those of the BTS. In addition, the leakage rate and the incidence of adjacent vertebral fractures after the HPDS treatment were reduced compared with those after treatment using the classic vertebroplasty devices. However, the BTS had better height and angle restoration abilities. CONCLUSIONS: The percutaneous HPDS with high-viscosity cement has similar clinical outcomes to those of traditional procedures in the treatment of vertebral fractures in the elderly. The HPDS with high-viscosity cement is better than the BTS in the treatment of mild and moderate OVCFs and could be an alternative method for the treatment of severe OVCFs.

Effect of leptin on long-term spatial memory of rats with white matter damage in developing brain / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the neuroprotective effect of leptin by observing its effect on spatial memory of rats with white matter damage in developing brain.</p><p><b>METHODS</b>A total of 80 neonatal rats were randomly divided into 3 groups: sham-operation (n=27), model (n=27) and leptin intervention (n=27). The rats in the model and leptin intervention groups were used to prepare a model of white matter damage in developing brain, and the rats in the leptin intervention group were given leptin (100 μg/kg) diluted with normal saline immediately after modelling for 4 consecutive days. The survival rate of the rats was observed and the change in body weight was monitored. When the rats reached the age of 21 days, the Morris water maze test was used to evaluate spatial memory.</p><p><b>RESULTS</b>There was no significant difference in the survival rate of rats between the three groups (P>0.05). Within 10 days after birth, the leptin intervention group had similar body weight as the sham-operation group and significantly lower body weight than the model group (P<0.05); more than 10 days after birth, the leptin intervention group had rapid growth with higher body weight than the model and sham-operation groups (P>0.05). The results of place navigation showed that from the second day of experiment, there was a significant difference in the latency period between the three groups (P<0.05); from the fourth day of experiment, the leptin intervention group had a similar latency period as the sham-operation and a significantly shorter latency period than the model group (P<0.05). The results of space search experiment showed that compared with the sham-operation group, the model group had a significant reduction in the number of platform crossings and a significantly longer latency period (P<0.05); compared with the model group, the leptin intervention group had a significantly increased number of platform crossings and a significantly shortened latency period (P<0.05), while there was no significant difference between the leptin intervention and sham-operation groups.</p><p><b>CONCLUSIONS</b>Leptin can alleviate spatial memory impairment of rats with white matter damage in developing brain. It thus exerts a neuroprotective effect, and is worthy of further research.</p>

Absorption, distribution, metabolism, excretion and toxicity of biologics and its application in pharmacokinetic modeling / 药学学报

Recently, more and more attentions of drug development are placed to macromolecules, such as monoclonal antibodies, proteins, etc. It has become one of the most promising areas in drug research and development in 21st Century. In terms of the structure and the ADMET (absorption, distribution, metabolism, excretion and toxicity), macromolecules is different from small molecule drugs, which lead to a distinct modeling strategy. The characterization of biologics ADMET processes and its application in the PK model selection of macromolecules are reviewed in this paper.

Computed tomography and magnetic resonance imaging features of pancreatic neuroendocrine tumors / 中华消化外科杂志

Objective To investigate the features of computed tomography (CT) and magnetic resonance imaging (MRI) on pancreatic neuroendocrine tumors (pNENs).Methods The retrospective and descriptive study was adopted.The clinicopathological data of 33 patients with pNENs who were admitted to the Tongji Hospital of Tongji Medical School of Huazhong University of Science and Technology between May 2012 and February 2016 were collected.All the patients underwent plain and enhanced scans of CT and MRI.Observation indicators:(1) overall imaging findings and pathological results of pNENs,(2) imaging findings of functional pNENs,(3) imaging findings of non-functional pNENs.Main analysis indicators included tumor diameter,location,boundary,density,cystic degeneration,enhancement,signal,calcification,with or without pancreaticobiliary duct dilation,with or without surrounding tissues invasion,lymph node and distant organ metastases.Results (1) Overall imaging findings and pathological results of pNENs:of 33 patinets with pNENs,24 underwent CT examination,3 underwent MRI examination and 6 underwent CT and MRI examinations.Tumors of 33 patients were solitary with a diameter of 0.6-16.0 cm.Ten,1,13 and 9 tumors were respectively located at the head of pancreas,uncinate process of pancreas,body of pancreas and tail of pancreas.Thirty-three patients were diagnosed as pNENs by pathological examination,including 20 with functional pNENs (insulinoma) and 13 with non-functional pNENs,and G1,G2 and G3 were respectively detected in 24,7 and 2 patients.The coincidence rate between preoperative CT or MRI examination and pathological examination was 90.9％ (30/33).One,1 and 1 patients were misdiagnosed as pancreatic cancer,enlargement of peripancreatic lymph nodes and duodenal gastrointestinal stromal tumor,respectively.(2) Imaging findings of functional pNENs:tumor diameter of 20 patients with functional pNENs was 0.6-3.0 cm with an average diameter of 1.5 cm.Fòur,10 and 6 tumors were respectively located at the head of pancreas,body of pancreas and tail of pancreas.Of 20 patients with functional pNENs,tumors of 19 patients showed clear boundary and 1 showed unclear boundary,and tumors of 18 patients had uniform density and 2 had uneven density with cystic degeneration,without the occurrence of calcification.Of 20 patients undergoing dynamic enhanced scans,tumors of 19 patients demonstrated obvious enhancement in arterial phase and slightly obvious enhancement or were equal to normal pancreatic tissues in portal vein phase and lag phase,and tumor of 1 patient demonstrated slight enhancement in arterial phase and was equal to or less than normal pancreatic tissues in portal vein phase and lag phase.Tumors in 3 patients undergoing MRI scans were manifested as hypointensity on T1-weighted imaging (T1WI),hyperintensity on T2WI and hyperintensity on DWI (b =1 000 s/m2),with clear imaging.Of 20 patients,1 was accompanied with atrophy of pancreatic tissues at distal tumor,pancreatic duct dilatation,multiple retention cyst and enlargement of lymph nodes around the hepatic artery.(3) Imaging findings of non-functional pNENs:tumor diameter of 13 patients with non-functional pNENs was 1.5-16.0 cm with an average diameter of 5.0 cm.Six,1,3 and 3 tumors were respectively located at the head of pancreas,uncinate process of pancreas,body of pancreas and tail of pancreas.Of 13 patients with non-functional pNENs,tumors of 11 patients showed clear boundary and 2 showed unclear boundary,tumors of 3 patients had uniform density and 10 had uneven density with cystic degeneration,and tumors of 2 patients had calcification.Of 13 patients undergoing dynamic enhanced scans,tumors of 12 patients demonstrated obvious enhancement in arterial phase,continuous enhancement in portal vein phase and lag phase and less obvious enhancement at cystic degeneration area,with marked enlargement of supplying arteries and draining veins in partial tumors.Tumor of 1 patient demonstrated slight enhancement,and its enhancement was slightly less than normal pancreatic tissues in arterial phase,portal vein phase and lag phase,with unclear boundary.Results of MRI scans in 6 patients showed that tumors of 4 patients were manifested as hypointensity on T1WI,slight hyperintensity or mixed signal on T2WI and hyperintensity on DWI (b =1 000 s/m2),and tumors of 2 patients were manifested as hypointensity on T1WI,hypointensity on T2WI and hyperintensity on DWI (b =800 s/m2).Of 13 patients with non-functional pNENs,4 had pancreaticobiliary duct dilation and 7 had local tissues invasion or distant organ metastasis (4 with liver metastasis,1 with peripanereatic lymph node metastasis,1 with liver and peripancreatic lymph node metastases and 1 with liver metastasis combined with splenic venous and arterial invasion),including 1 in G1,4 in G2 and 2 in G3.Of 5 patients with tumor diameter ＞ 5.0 cm,4 were complicated with liver or lymph node metastases.Conclusions CT and MRI features of pNENs have a certain characteristics.For functional pNENs,benign and solid tumor is common,with clear boundary and smaller diameter.For non-functional pNENs,tumor size is bigger and cystic necrosis occurs within the tumor,with various enhancements.

Isolation and analysis of differentially expressed genes from peanut in response to challenge with Ralstonia solanacearum

Background: Bacterial wilt caused by Ralstonia solanacearum is the most devastating disease in peanut. Planting resistant peanut cultivars is deemed as the sole economically viable means for effective control of the disease. To understand the molecular mechanism underlying resistance and facilitate breeding process, differences in gene expression between seeds of Rihua 1 (a Virginia type peanut variety resistant to bacterial wilt) inoculated with the bacterial pathogen suspension (10(9) cfu ml-1) and seeds of the same cultivar treated with water (control), were studied using the GenefishingTM technology. Results: A total of 25 differentially expressed genes were isolated. Expression of genes encoding cyclophilin and ADP-ribosylation factor, respectively, were further studied by real time RT-PCR, and full length cDNAs of both genes were obtained by rapid amplification of cDNA ends. Conclusions: The study provided candidate genes potentially useful for breeding peanut cultivars with both high yield and bacterial wilt resistance, although confirmation of their functions through transgenic studies is still needed.

Effect of marine collagen peptides on markers of metabolic nuclear receptors in type 2 diabetic patients with/without hypertension / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To explore Effects of marine collagen peptides (MCPs) on markers of metablic nuclear receptors, i.e peroxisome proliferator-activated receptor (PPARs), liver X receptor (LXRs) and farnesoid X receptor (FXRs) in type 2 diabetic patients with/without hypertension. METHOD Study population consisted of 200 type 2 diabetic patients with/without hypertension and 50 healthy subjects, all of whom were randomly assigned to MCPs-treated diabetics (n = 50), placebo-treated diabetics (n = 50), MCPs-treated diabetics with hypertension (n=50), placebo-treated diabetics with hypertension (n = 50), and healthy controls (n = 50). MCPs or placebo (water-soluble starch) were given daily before breakfast and bedtime over three months. Levels of free fatty acid, cytochrome P450, leptin, resistin, adiponectin, bradykinin, NO, and Prostacyclin were determined before intervention, and 1.5 months, and 3 months after intervention. Hypoglycemia and the endpoint events during the study were recorded and compared among the study groups.</p><p><b>RESULT</b>At the end of the study period, MCPs-treated patients showed marked improvement compared with patients receiving placebo. The protection exerted by MCPs seemed more profound in diabetics than in diabetics with hypertension. In particular, after MCPs intervention, levels of free fatty acid, hs-CRP, resistin, Prostacyclin decreased significantly in diabetics and tended to decrease in diabetic and hypertensive patients whereas levels of cytochrome P450, leptin, NO tended to decrease in diabetics with/without hypertension. Meanwhile, levels of adiponectin and bradykinin rose markedly in diabetics following MCPs administration.</p><p><b>CONCLUSION</b>MCPs could offer protection against diabetes and hypertension by affecting levels of molecules involved in diabetic and hypertensive pathogenesis. Regulation on metabolic nuclear receptors by MCPs may be the possible underlying mechanism for its observed effects in the study. Further study into its action may shed light on development of new drugs based on bioactive peptides from marine sources.</p>

Etiology analysis of 38 patients with hemophagocytic syndrome / 中国实验血液学杂志

The study was aimed to investigate the etiology and the clinical characteristics of patients with hemophagocytic syndrome. The clinical data of 38 patients with hemophagocytic syndrome were retrospectively analyzed, and prf1 and stx11 were detected for the mutational analysis. The results showed that 38 cases were diagnosed as hemophagocytic syndrome, including 1 case of familial hemophagocytic lymphohistiocytosis (FHL), 14 cases associated with infectious disease (36.84%), 10 cases with malignancies (26.32%), 7 cases with rheumatic disease (18.42%), other 6 cases of unknown etiology (15.79%). 9 out of 38 cases died with mortality of 23.68%, including 4 cases associated with infectious disease, 2 cases with malignancies, 1 case with rheumatic disease, and 2 cases of unknown etiology. One case was found to have prf1 mutation, and was diagnosed as FHL at last. It is concluded that the causes of HPS are diverse, different etiology results in different outcome. It is important to find etiology when HPS is diagnosed, and prf1 and stx11 genetic analysis plays a important role in the diagnosis of FHL.

Transforming growth factor beta alleviates acute graft-versus-host-disease after allogeneic bone marrow transplantation in murine model / 中国实验血液学杂志

This study was purposed to investigate the effects and mechanism of transforming growth factor beta (TGF-beta) on acute graft-versus-host disease (aGVHD) after allogeneic bone marrow transplantation (allo-BMT). The recipients were male BABL/c mice, while the donors were male C57BL/6 mice. The murine model of aGVHD had been established by allo-BMT with donor derived T cells. Experiment was divided into four groups: control group, radiation control group, transplantation control group and TGF-beta treated group. Mice in TGF-beta treated group were daily subcutaneously injected TGF-beta1 (1 microg/kg) in two days before transplantation until seven days after it. The results showed that the survival time of mice in TGF-beta treated group was significantly longer than that in transplantation control group, and the aGVHD pathological changes in TGF-beta treated group were milder than that in transplantation control group. At seven days after transplantation, the level of IL-2 in TGF-beta treated group was significantly higher than that in control group, but significantly lower than that in transplantation control group. The level of IL-10 in TGF-beta treated group was significantly higher than that in transplantation control group, but the level of IL-10 in transplantation control group was significantly lower than that in other groups. It is concluded that TGF-beta may alleviate or suppress lethal aGVHD, and elevate the survival rate after allo-BMT in murine model. Accommodating of the Th1 and Th2 cytokine levels is the possible mechanism of TGF-beta preventing lethal aGVHD.

Diagnostic significance of NK cell activity and soluble CD25 level in serum from patients with secondary hemophagocytic lymphohistiocytosis / 中国实验血液学杂志

This study was aimed to explore the clinical significance of NK cell activity and serum soluble CD25 (sCD25) level in early diagnosis of the patients with secondary hemophagocytic lymphohistiocytosis (HLH). 38 suspected secondary HLH patients from June 2005 to June 2008 and 25 healthy subjects were enrolled in the study. The NK cell activity in peripheral blood was determined by a released LDH assay, The sCD25 level in serum was detected with ELISA double antibody sandwich assay. The 38 suspected secondary HLH patients were divided into diagnosed and excluded group according to HLH-2004 diagnostic criteria, The NK cell activity and sCD25 level were compared between the two groups. The results showed that 22 out of 38 suspected patients were diagnosed as secondary HLH, the NK cell activity in peripheral blood of these 22 patients was significantly lower than that of healthy control (p < 0.001), the sCD25 level in peripheral blood of these 22 diagnosed patients was higher than that of healthy control (p < 0.05). In conclusion, detection of NK cell activity and sCD25 level may be valuable in the early diagnosis of secondary HLH.

Diagnostic significance of glycosylated ferritin for patients with secondary hemophagocytic lymphohistiocytosis / 中国实验血液学杂志

This study was aimed to explore the level of glycosylated ferritin in the patients with secondary hemophagocytic lymphohistiocytosis (HLH) and its diagnostic significance. 29 suspected HLH patients from October 2007 to October 2008 were enrolled in the study, and 25 healthy subjects were selected as control. The 29 suspected HLH patients were divided into confirmed group (22 out of 29) and unconfirmed group (7 out of 29) according to HLH-2004 diagnostic criteria. The percentage of glycosylated ferritin in peripheral blood was determined by phytohemagglutinin adsorption assay. The results showed that the median level of total serum ferritin in patients of confirmed group (2897.6+/-1837.2 microg/L) was significantly higher than that in patients of unconfirmed group (653.1+/-249.1 microg/L) (p<0.01), and was also higher than that in controls (414.6+/-212.6 microg/L) (p<0.01). The median percentage of glycosylated ferritin in patients of confirmed group was significantly lower (17.0+/-4.2%) than that in patients of unconfirmed group (40.7+/-4.5%) (p<0.01) and was lower than that in controls (53.6+/-13.3%) (p<0.01). The sensitivity (86.4% vs 77.3%) and specificity (71.4% vs 42.9%) of glycosylated ferritin for the diagnosis of HLH were higher than that of total serum ferritin. In conclusions, glycosylated ferritin may be a helpful marker for the diagnosis of HLH.

Diagnostic significance of NK cell activity in the patients with hemophagocytic syndrome / 中国实验血液学杂志

This study was aimed to explore the level of NK cell activity in the patients with secondary hemophagocytic syndrome (HPS) and its significance for early diagnosis of this disease. 16 suspected HPS patients and 25 healthy subjects were enrolled in this study. The activity of NK cells in peripheral blood was detected by a released LDH assay. The activity level of NK cells in peripheral blood from patients who were finally diagnosed as HPS was compared with healthy subjects. The results showed that 8 out of 16 suspected HPS patients were finally diagnosed as secondary HPS. The activity of NK cells in peripheral blood of these 8 patients was obviously lower than that in healthy subjects with statistical significance (p<0.001), and showed abnormal in the early stage of this disease. It is concluded that the detection of NK cell activity may play an important role in earlier diagnosis of secondary hemophagocytic syndrome.