Blood diagnostic and prognostic biomarkers in amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis is a devastating neurodegenerative disease for which the current treatment approaches remain severely limited.The principal pathological alterations of the disease include the selective degeneration of motor neurons in the brain,brainstem,and spinal cord,as well as abnormal protein deposition in the cytoplasm of neurons and glial cells.The biological markers under extensive scrutiny are predominantly located in the cerebrospinal fluid,blood,and even urine.Among these biomarke rs,neurofilament proteins and glial fibrillary acidic protein most accurately reflect the pathologic changes in the central nervous system,while creatinine and creatine kinase mainly indicate pathological alterations in the peripheral nerves and muscles.Neurofilament light chain levels serve as an indicator of neuronal axonal injury that remain stable throughout disease progression and are a promising diagnostic and prognostic biomarker with high specificity and sensitivity.However,there are challenges in using neurofilament light chain to diffe rentiate amyotrophic lateral sclerosis from other central nervous system diseases with axonal injury.Glial fibrillary acidic protein predominantly reflects the degree of neuronal demyelination and is linked to non-motor symptoms of amyotrophic lateral sclerosis such as cognitive impairment,oxygen saturation,and the glomerular filtration rate.TAR DNA-binding protein 43,a pathological protein associated with amyotrophic lateral sclerosis,is emerging as a promising biomarker,particularly with advancements in exosome-related research.Evidence is currently lacking for the value of creatinine and creatine kinase as diagnostic markers;however,they show potential in predicting disease prognosis.Despite the vigorous progress made in the identification of amyotrophic lateral sclerosis biomarkers in recent years,the quest for definitive diagnostic and prognostic biomarke rs remains a formidable challenge.This review summarizes the latest research achievements concerning blood biomarkers in amyotrophic lateral sclerosis that can provide a more direct basis for the differential diagnosis and prognostic assessment of the disease beyond a reliance on clinical manifestations and electromyography findings.

Study on Polymorphisms in the Blood Protein of Tibetan Mastiff

[ Objective] The aim was to study the protein polymorphism in the blood of Tibetan Mastiff, and provide some theoretical basis for resource protection and reasonable development and utilization of Tibetan Mastiff varieties. [ Method] A total of 103 blood samples were taken from four populations of Hequ Tibetan Mastiff, Qinhai Tibetan Mastiff, Tibetan Spaniel and native dogs of Qinghai. Seven blood protein Iocus（Tf, Po, Sα2, Hb, AIb, Pr and Amy）were investigated by using vertical polyacrylamide gel electrophoresis with discontinuous buffer system. Then the genetic variation during different populations was analyzed. [ Result] Genetic variations were observed in Tf, Sα2 and Po in four populations, others were not polymorphic. There were three alleles at the locus of Tf and Po, two alleles at the loci of Sα2. Effective number of alleles and Nei＇s average expected heterozygosity were 1. 532 4 and 0.230 3 relatively, all higher in Tibetan Mastiff than other populations. [ Conclusion] Protein locus in blood of Tibetan Mastiff existed in genetic variation.

Effect of Wheat Based Diet Supplemented with Xylanase on Blood Sugar and Total Protein in Serum of Geese

[Objective] The effects of wheat based diet supplemented with xylanase on blood sugar and total protein in serum of geese were studied. [ Method ] By using the randomized design of single factor, the 1-day-old healthy goslings were divided into 6 groups and fed with corn based diet, wheat based diet and wheat based diet supplemented with xylanase at different concentrations respectively, the contents of blood sugar and total protein in serum were determined. [ Result] The wheat based diet supplemented with xylanase could increase the blood sugar and total protein in serum of geese and wheat based diet supplemented with 0.2% xylanase generated the best effect, which was higher than those of corn based diet group. As for the concentration of protein in serum, wheat based diet supplemented with 0.2% xylanase was significantly different from corn based diet and wheat based diet. [ Conclusion] The wheat based diet supplemented with xylanase could enhance geese production.

Long-term effects of early antibiotic intervention on blood parameters,apparent nutrient digestibility, and fecal microbial fermentation profile in pigs with different dietary protein levels

Backgroud： This study aimed to determine the effects of early antibiotic intervention（EAI） on subsequent blood parameters, apparent nutrient digestibility, and fecal fermentation profile in pigs with different dietary crude protein（CP） levels. Eighteen litters of piglets（total 212） were randomly allocated to 2 groups and were fed a creep feed diet with or without in-feed antibiotics（olaquindox, oxytetracycline calcium and kitasamycin） from postnatal d 7 to d 42. On d 42, the piglets within the control or antibiotic group were mixed, respectively, and then further randomly assigned to a normal-（20%, 18%, and 14% CP from d 42 to d 77, d 77 to d 120, and d 120 to d 185,respectively） or a low-CP diet（16%, 14%, and 10% CP from d 42 to d 77, d 77 to d 120, and d 120 to d 185,respectively）, generating 4 groups. On d 77（short-term） and d 185（long-term）, serum and fecal samples were obtained for blood parameters, microbial composition and microbial metabolism analysis.Results： EAI increased（P 〈 0.05） albumin and glucose concentrations in low-CP diet on d 77, and increased（P 〈 0.05） urea concentration in normal-CP diet. On d 185, EAI increased（P 〈 0.05） globulin concentration in normal-CP diets, but decreased glucose concentration. For nutrient digestibility, EAI increased（P 〈 0.05）digestibility of CP on d 77. For fecal microbiota, the EAI as well as low-CP diet decreased（P 〈 0.05） E. coli count on d 77. For fecal metabolites, on d 77, EAI decreased（P 〈 0.05） total amines concentration but increased skatole concentration in low-CP diet. On d 185, the EAI increased（P 〈 0.05） putrescine and total amines concentrations in low-CP diets but reduced（P 〈 0.05） in the normal-CP diets. The low-CP diet decreased the concentrations of these compounds.Conclusions： Collectively, these results indicate that EAI has short-term effects on the blood parameters and fecal microbial fermentation profile. The effects of EAI varied between CP levels, which was characterized by the significant alteration of glucose and putrescine concentration.

Productive performance and blood metabolites as affected by protected protein in sheep

This investigation included two experiments. Experiment 1 was executed to study the effect of feeding different rations of protected protein of canola meal on digestibility and nutritive values within sheep. Twenty male, healthy sheep were divided into five treatments according to the methods of protein protection (control, heat, sodium hydroxide, formaldehyde, and acetic acid treatments). Experiment 2 was carried out on developing lambs to investigate the effect of protected protein on growth performance and some blood metabolites. Animals in this ex-periment were also divided into the same treatments as Experiment 1. Animals in the first and second experiment were fed concentrate ration (80%) and wheat straw (20%) to cover the feed requirements. Nutritive values expressed as total digestible nutrients (TDN %) and digestible crude protein (DCP%) of the experimental rations was calculated. In the second experimental all animals were weighed biweekly and the amounts of rations were adjusted throughout the experimental period (120 days) according to their body weight change. Results indicated that in the first experimental protected protein by heat (HE) and sodium hydroxide (NH) had positive (P<0.05) effects on most of digestibility coefficients of different nutrients. Protein protection methods also improved (P<0.05) the nutritive values (TDN and DCP) in the HE treatment and NH treatment. In the second experiment body weight increased by 14% and 7% and also daily gain by 27% and14 % in HE and NH, respectively, while FM and AC decreased body weight by 8% and 4.4%. Higher values (P<0.01) in both thyroid hormones were observed in HE and NH than those other treatments. Also, higher values (P<0.01) of total protein, albumin, and glucose were observed in HE and NH than other treatments. The control (CTL) group recorded higher concentrations of urea-N and creatinine at different periods of the experiment in comparison with other treatments. Generally, from the present investigation it can be concluded that protected protein of canola meal by heat or sodium hydroxide treatments were more efficient for productive performance and some blood metabolites of sheep.

Correlation between intakes of carbohydrates,protein,and fat with random blood sugar levels in menopausal women

Objective: Carbohydrates, proteins, and fats are energy sources needed by the body for performing daily activities and generating primary energy substances. In women who have undergone menopause, the function of thyroid hormone in their body begins to decline and thus affects the ability of the body to produce energy. The purpose of this study was to determine the correlation between the intakes of carbohydrates, proteins, and fats and random blood sugar levels in menopausal women.Methods: This study was a correlational analytical research with a cross-sectional design, which was conducted in 72 menopausal women recruited by the purposive sampling technique. In this study, a 24-hour food recall form was used, and randomized blood sugar levels were measured using a glucometer. The analysis of the data was performed using a Pearson product moment and multiple linear regression.Results: Carbohydrates and fats together affected random blood sugar levels with an F-value of 25.810 and a p-value of 0.000.Meanwhile, adjusted R^2 showed the value of 0.411, indicating that the difference in the intake of carbohydrates and fats together affected random blood sugar levels by 41.1%.Conclusions: Intake of carbohydrates and fats affected random blood sugar levels, whereas the rest were influenced by other factors,and protein intake was unrelated to random blood sugar levels in menopausal women.

Protective role of buffalo pineal proteins on arsenic-induced oxidative stress in blood and kidney of rats

Objective: Exposure to various toxic metals has become an increasingly recognized source of ill- ness in human and animals, worldwide. Arsenic (As) and its compounds cause adverse health effects in animals and humans. Recently, it has been suggested that the pineal gland may also have antioxidants role due to secretary product other than melatonin. With keeping this view, pre-sent investigation tested effect of buffalo (Bubalus bubalis) pineal proteins (PP) on arsenic-induced oxidative stress in RBCs (Red blood cells) and kidney of rats. Methods: Eighteen adult female Wistar rats were grouped into group-I (Control), group-II (Arsenic control), and group-III (Arsenic + Pineal proteins). Experimental rats were given 100 ppm arsenic (p.o.) for 4 weeks alone or along with pineal proteins at a dose of 100 μg/kg body weight (i.p.). Results: Interestingly, arsenic ex-posure led to the stimulation of kidney catalase (CAT) activity, but inhibition of RBCs CAT activ-ity and significantly (P<0.05) increased the RBCs and kidney lipid peroxidation level (LPO). How-ever, arsenic treatment caused depletion of glu- tathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR) in kidney tissues. In RBCs, only GR and CAT activity were significantly (P<0.05) declined. These changes were significantly (P<0.05) reversed by PP treatment in arsenic exposed animals. Conclusion: Therefore, present study indicated the significant protecting effect of buf-falo (Bubalus bubalis) PP against arsenic in-duced-oxidative stress through antioxidant de-fense systems in rats.

Mechanism of hyperproteinemia-induced blood cell homeostasis imbalance in an animal model

Hyperproteinemia is a metabolic disorder associated with increased plasma protein concentration(PPC)and is often clinically complicated by malignant diseases or severe infections.At present,however,research on the molecular mechanism underlying high PPC(HPPC)is scant.Here,an animal model of primary hyperproteinemia was constructed in an invertebrate(Bombyx mori)to investigate the effects of HPPC on circulating blood cells.Results showed that HPPC affected blood cell homeostasis,leading to increased reactive oxygen species levels,and induced programmed cell death dependent on the endoplasmic reticulum-calcium ion signaling pathway.HPPC induced the proliferation of blood cells,mainly granulocytes,by activating the Janus kinase/signal transducer and activator of transcription(JAK/STAT)signaling pathway.Supplementation with the endocrine hormone active substance 20 E significantly reduced the impact of HPPC on blood cell homeostasis.Thus,we identified a novel signaling pathway by which HPPC affects blood cell homeostasis,which differs from hyperglycemia,hyperlipidemia,and hypercholesterolemia.In addition,we showed that down-regulation of gene expression of the hematopoietic factor Gcm could be used as a potential early detection indicator for hyperproteinemia.

Effect of Xuesaitong Combined with Autologous Platelet-rich Gel on Blood Glucose,AT-III,TNF-αand 24-h Urinary Protein Level in Patients with Diabetic Foot

[Objectives]To investigate the clinical effect of different doses of Xuesaitong combined with autologous platelet-rich gel(APG)on patients with diabetic foot(DF).[Methods]90 patients with diabetic foot admitted to our hospital from February 2017 to February 2019 were enrolled in the study.According to the random number table method,the subjects were divided into study group A and B and control group C.Group A was given a low dose of Xuesaitong combined with APG,while group B was given high-dose Xuesaitong combined with APG and group C was treated only with APG.Patients in the three groups were observed and the changes of related indexes were detected.[Results]After treatment,with regard to the three groups,the fasting blood glucose,2-h postprandial blood glucose level,HbA1c,TNF-α,Hcy,blood urea nitrogen(BUN),creatinine(Cr)and 24-h urine protein levels were all decreased,yet AT-III level was increased,and granulation tissue coverage and thickness,wound clearance rate were increased,while the repair time of ulcer surface was significantly reduced.The above indexes were all significantly different(P<0.05).Compared with the control group C,the changes of the indexes in the study group A and B were the same as above,and the difference was significant(P<0.05).Compared with the study group A,the level of AT-III in group B was significantly increased(P<0.05),while TNF-α,Hcy,BUN,Cr and 24-h urine protein levels were significantly decreased(P<0.05).There was no significant difference in the other indexes between groups A and B(P>0.05).[Conclusions]Xuesaitong combined with APG could effectively reduce the blood sugar level of DF patients,improve the clinical indexes,promote wound healing,and the high-dose group had more significant advantages and was worthy of promotion.

Transactivating-transduction protein-polyethylene glycol modified liposomes traverse the blood-spinal cord and blood-brain barriers

Naive liposomes can cross the blood-brain barrier and blood-spinal cord barrier in small amounts. Liposomes modified by a transactivating-transduction protein can deliver antibiotics for the treatment of acute bacterial infection-induced brain inflammation. Liposomes conjugated with polyethylene glycol have the capability of long-term circulation. In this study we prepared transactivating-transduction protein-polyethylene glycol-modified liposomes labeled with fiuorescein isothiocyanate. Thus, liposomes were characterized by transmembrane, long-term circulation and fluorescence tracing. Uptake, cytotoxicity, and the ability of traversing blood-spinal cord and blood-brain barriers were observed following coculture with human breast adenocarcinoma cells （MCF-7）. Results demonstrated that the liposomes had good biocompatibility, and low cytotoxicity when cocultured with human breast adenocarcinoma cells. Liposomes could traverse cell membranes and entered the central nervous system and neurocytes through the blood-spinal cord and blood-brain barriers of rats via the systemic circulation. These results verified that fluorescein isothiocyanate-modified transactivating-transduction protein-polyethylene glycol liposomes have the ability to traverse the blood-spinal cord and blood-brain barriers.

The effects of smoking on blood lipids, C-reactive protein, and homocysteine in young patients with ischemic stroke

Objective:To investigate clinical significance of the effects of smoking on blood lipids, C-reactive protein,and homocysteine in young ischemic stroke patients.Methods:The clinical data of 423 young stroke patients in the department of neurology at Taihe Hospital in Shiyan City, China were retrospectively analyzed, including age,gender,drinking history,family history,and atrial fibrillation history. Patients were divided into two groups according to whether they smoked,and the blood lipids, C-reactive protein, and homocysteine were compared between groups.Results:The proportion of smokers was 41.83%.The levels of total cholesterol,triglycerides (TG), low density lipoprotein (LDL), and homocysteine were higher in patients who smoked than in those who did not(P < 0.05). High density lipoprotein (HDL) was lower in the smoking group (P < 0.05). C-reactive protein test results were divided into groups according to whether the levels exceeded the normal range or not, and no correlation was found between C-reactive protein levels and smoking(P>0.05). Conclusion:Total cholesterol, TG, LDL, HDL, and homocysteine were significantly different between stroke patients who smoked and those who did not. We therefore suggest that smoking cessation take place as soon as possible and that it be avoided entirely in order to reduce the incidence of atherosclerosis and stroke.

Comparison of the level of thrombus precursor protein in blood plasma between patients with acute cerebral infarction and healthy persons at different time

BACKGROUND： Thrombus precursor protein （TpP） is the index of thrombus activity level, and it is also early referencing index in detecting thrombus diseases. OBJECTIVE： To dynamically observe the changes of TpP level in blood plasma of patients with acute cerebral infarction at different time after onset, and to compare the differences of plasma TpP level between patients with acute cerebral infarction and healthy persons who received health examination. DESIGN： Controlled observation SETTING： Department of Neurology, Affiliated Hospital of Xuzhou Medical College PARTICIPANTS： Totally 58 patients with acute cerebral infarction who received the treatment in the Department of Neurology, Affiliated Hospital of Xuzhou Medical College between September 2004 and March 2005 were recruited in this study. They all met the diagnostic criteria revised by the 4^th National Conference of Cerebrovascular Disorders in 1995 and were diagnosed by clinical and skull CT and （or） MRI examinations. The patients included 33 male and 25 female aged from 36 to 87 years. Time to onset 〈 6 hours, 6 to 11 hours, 12 to 23 hours, 24 to 48 hours and 〉 48 hours were found in 10,11,14,10 and 13 patients respectively. Another 51 persons who homeochronously received the health body examination in our hospital were recruited, including 34 male and 17 female, aged 38 to 85 years, serving as control group. Patients with cardio-cerebrovascualr diseases or liver and kidney diseases were excluded. All the involved subjects were informed of the detected items. METHODS： About 4 mL venous blood was respectively taken from patients admitted to the hospital within 6 hours, 6 toll hours, 12 to 23 hours, 24 to 48 hours and more then 48 hours after onset, and healthy persons when receiving health examination. The level of TpP in blood plasma was measured with enzymelinked immunosorbent assay. MAIN OUTCOME MEASURES： ① Comparison of the level of plasma TpP between patients and controls;② Comparison of the level of plasma TpP of patients with acute cerebral infarction at different time after onset. RESULTS： Totally 58 patients with acute cerebral infarction and 51 persons who received health examination participated in the result analysis. ①Comparison of plasma TpP level between patients and controls： The plasma TpP level of patients with acute cerebral infarction was significantly higher than that of control group [（16.12±3.28）vs （5.38±1.36） mg/L, t= 20.993, P〈 0.01 ]. ② Comparison of plasma TpP level of patients with acute cerebral infarction at different time after onset： The level of plasma TpP was （12.06±3.06） mg/L within 6 hours, （15.11±3.42） mg/L at 6 to 11 hours, （20.63±4.05） mg/L at 12 to 23 hours, （16.15±3.50） mg/L at 24 to 48 hours and （11.88±3.11） mg/L at more than 48 hours after onset. It increased from the 6^th hour, reached the peak at the 12^th to 23^rd hours, maintained at very high level at the 48= hour and then gradually decreased and recovered to the level within 6 hours after onset. The level of plasma TpP of patients with acute cerebral infarction was signiticantly higher at the 12^th to 23^rd hours after onset and the 24^th to 48^th hours after onset than within 6 hours after onset （t = 13.385, P 〈 0.05）. CONCLUSION： ①The level of plasma TpP of patients with acute cerebral infarction is significantly higher than that of persons who received health examination.② Plasma TpP levels of patients with acute cerebral infarction change in wave manner at the different time after onset.

Monitoring the changes in plasm C-reactive protein,fibrinogen and blood white cell in patients with primary hypertension combined with acute cerebral infarction

BACKGROUND： Inflammatory reaction and the increased level of its accompanying active protein play an important role in the occurrence and development of cerebral infarction. C-reactive protein, fibrinogen and white blood cell, as the monitoring index of inflammatory reaction, are very important in the occurrence and development of acute cerebral infarction. OBJECTIVE： To make a comparison between patients with primary hypertension accompanied with acute cerebral infarction and with simple primary hypertension by observing the changes in plasma C-reactive protein and fibrinogen levels as well as white blood cell and differential counts and analyzing their significances. DESIGN ： Controlled observation SETTING ： Ward Building for VIP, Shenzhen Hospital, Peking University. PARTICIPANTS： Totally 133 patients with primary hypertension were selected from Ward Building for VIP, Shenzhen Hospital, Peking University during September 2003 to September 2005, The diagnostic criteria were based on the hypertension diagnosis criteria formulated by the 7^th World Health Organization-International Society of Hypertension Guidelines （WHO-ISH） in 1998. The informed consents were obtained from all the participants. The involved patients were assigned into two groups： primary hypertension group, in which, there were 65 patients with primary hypertension （ degree 2）, including 42 males and 23 females, with mean age of （61 ±14）years and mean blood pressure of （162.7±6.8）/（94.2±8.4） mm Hg （1 mm Hg =0.133 kPa）, and primary hypertension combined with cerebral infarction group, in which, there were 68 patients with primary hypertension combined with cerebral infarction （ meeting the diagnostic criteria formulated in the 4^th National Cerebrovascular Diseases Meeting in 1995 and diagnosed by skull CT or MRI to exclude the patients with lacunar infarction）, including 42 males and 26 females, with mean age of （56±15） years and mean blood pressure of （176.4±9.2）/（96.3±9.7） mm Hg. METHODS： Plasm C-reactive protein and fibrinogen levels, and white blood cell and differential counts of patients in the two groups were examined 24 hours after stroke. The above indexes were re-examined in the primary hypertension combined with cerebral infarction group 72 hours after stroke. White blood cell and differential counts were performed with laser method （East Asia FE-95001 RAM-1, Japan）. The level of C-reactive protein was measured with turbidimetry （BNII Automatic Systems For Analysis, USA）. The level of fibrinogen was measured with algorithm method when prothrombin time was normal and with Clauss method when prothrombin time was abnormal （ACL Automatic Coagulation Analyzer, USA）. MAIN OUTCOME MEASURES： The plasm C-reactive protein and flbrinogen levels, and white blood cell and differential counts 24 hours after stroke in two groups and 72 hours after stroke in primary hypertension combined with cerebral infarction group. RESULTS： All the 133 involved patients participated in the result analysis. The plasm C-reactive protein and fibrinogen levels, and white blood cell and neutrophil counts in patients with primary hypertension were all within the normal range. The plasm C-reactive protein and fibrinogen levels, and white blood cell and neu- trophil counts in patients with primary hypertension combined with cerebral infarction were significantly higher than those in patients with primary hypertension 24 hours after stroke and 72 hours after stroke respectively[24 hours after stroke：（32.12±11.76） mg/L vs. （5.02±3.21 ） mg/L;（4.64±0.75） g/L vs. （3.12±0.49） g/L; （9.32±81）×10^9 L^- 1 vs. （5.78±1.32）×10^9L^- 1 （7.85±2.38）×10^9 L^- 1 vs.（3.49±1.28）×10^9 L^-1,t =7.094, 5.759,4.106,5.491, respectively,all P〈 0.01; 72 hours after stroke： （47.62±18.43） mg/L vs. （32.12±11.76） mg/L; （5.08±0.82） g/L vs. （4.64±0.75） g/L, t =2.864,2.220, respectively, both P 〈 0.05]. CONCLUSION： The increase in fibrinogen level and white blood cell count are the important index in monitoring primary hypertension combined with acute cerebral infarction. The increase in plasm C-reactive protein and fibrinogen levels 72 hours after stroke indicates that plasma C-reactive protein and fibrinogen are very important in the development of disease.

Relationship of gelatinases-tight junction proteins and blood-brain barrier permeability in the early stage of cerebral ischemia and reperfusion

Gelatinases matrix metalloproteinase-2 and matrix metalloproteinase-9 have been shown to mediate claudin-5 and occludin degradation, and play an important regulatory role in blood-brain barrier permeability. This study established a rat model of 1.5-hour middle cerebral artery occlusion with reperfusion. Protein expression levels of claudin-5 and occludin gradually decreased in the early stage of reperfusion, which corresponded to the increase of the gelatinolytic activity of matrix metalloproteinase-2 and matrix metalloproteinase-9. In addition, rats that received treatment with matrix metalloproteinase inhibitor N-[（2R）-2-（hydroxamidocarbonylmethyl）-4-methylpenthanoyl]-L- tryptophan methylamide （GM6001） showed a significant reduction in Evans blue leakage and an inhibition of claudin-5 and occludin protein degradation in striatal tissue. These data indicate that matrix metalloproteinase-2 and matrix metalloproteinase-9-mediated claudin-5 and occludin degradation is an important reason for blood-brain barrier leakage in the early stage of reperfusion. The leakage of the blood-brain barrier was present due to gelatinases-mediated degradation of claudin-5 and occludin proteins. We hypothesized that the timely closure of the structural component of the blood-brain barrier （tight junction proteins） is of importance.

Relationship between the expression of IP-10 and IP-10 mRNA in peripheral blood and HBV DNA level in patients with cirrhosis

BACKGROUND: Post-hepatitic cirrhosis is regarded as common and severe form of liver damage. Interferon gamma-inducible protein 10 (IP-10), a member of the non-ELR (glutamic-leucine-arginine) motif CXC chemokine family, has recently been shown to recruit and activate specific subsets of leukocytes to sites of inflammation or an immune response during the development of hepatic cirrhosis. However, the effects of IP-10 and IP-10 mRNA on inflammatory infiltration at local sites and in the peripheral blood of patients with post-hepatitic cirrhosis as well as their relationship with viral load are still poorly defined. This study aimed to detect the relationship between the expression of IP-10 in serum, IP-10 mRNA in peripheral blood mononuclear cells (PBMCs), and the levels of HBV DNA in the serum of patients, and to explore their role in the pathogenesis of cirrhosis. METHODS: Typical patients with cirrhosis after HBV infection were selected, and their serum IP-10 concentrations were evaluated with ELISA, the content of IP-10 mRNA in PBMCs was measured by real-time PCR, and the load of HBV DNA in serum and PBMCs was assessed by semi-quantitative analysis of gel imaging. RESULTS: The levels of IP-10 in serum and IP-10 mRNA in PBMCs of patients with cirrhosis were 299.9 +/- 77.2 pg/ml and 0.7500 +/- 0.1495, respectively. They were higher than those of controls (P<0.05) and also increased in the HBV DNA(+) groups (P<0.05, P<0.01) to 343.0 +/- 80.3 pg/ml and 0.8465 +/- 0.1528, respectively. The levels of IP-10 in serum and IP-10 mRNA in PBMCs were clearly correlated with the load of HBV DNA (P<0.01). CONCLUSIONS: The levels of IP-10 and IP-10 mRNA in the peripheral blood of patients with cirrhosis increase are closely correlated with the load of HBV DNA in serum, and play a key role in the progression of post-hepatitic cirrhosis. (Hepatobiliary Pancreat Dis Int 2010; 9: 280-286)

Correlation of fibrinogen-like protein 2 with progression of acute pancreatitis in rats

AIM: To examine fibrinogen-like protein 2 (fgl2) expression during taurocholate-induced acute pancreatitis progression in rats and its correlation with pancreatic injury severity. METHODS: Forty-eight male Sprague-Dawley rats were randomly divided into the severe acute pancreatitis (SAP) group (n = 24) and the sham operation (SO) group (n = 24). Sodium taurocholate (4% at doses of 1 mL/kg body weight) was retrogradely injected into the biliopancreatic ducts of the rats to induce SAP. Pancreatic tissues were prepared immediately after sacrifice. At the time of sacrifice, blood was obtained for determination of serum amylase activity and isolation of peripheral blood mononuclear cells (PBMCs). Pancreatic tissue specimens were obtained for routine light microscopy including hematoxylin and eosin staining, and the severity of pancreatic injury was evaluated 1, 4 and 8 h after induction. Expression of fgl2 mRNA was measured in the pancreas and PBMCs using reverse transcription polymerase chain reaction. Expression of fgl2 protein was evaluated in pancreatic tissues using Western blotting and immunohistochemical staining. Masson staining was also performed to observe microthrombosis. RESULTS: At each time point, levels of fgl2 mRNAs in pancreatic tissues and PBMCs were higher (P < 0.05) in the SAP group than in the SO group. For pancreatic tissue in SAP vs SO, the levels were: after 1 h, 3.911 ± 1.277 vs 1.000 ± 0.673; after 4 h, 9.850 ± 3.095 vs 1.136 ± 0.609; and after 8 h, 12.870 ± 3.046 vs 1.177 ± 0.458. For PBMCs in SAP vs SO, the levels were: after 1 h, 2.678 ± 1.509 vs 1.000 ± 0.965; after 4 h, 6.922 ± 1.984 vs 1.051 ± 0.781; and after 8 h, 13.533 ± 6.575 vs 1.306 ± 1.179. Levels of fgl2 protein expression as determined by Western blotting and immunohistochemical staining were markedly up-regulated (P < 0.001) in the SAP group compared with those in the SO group. For Western blotting in SAP vs SO, the results were: after 1 h, 2.183 ± 0.115 vs 1.110 ± 0.158; after 4 h, 2.697 ± 0.090 vs 0.947 ± 0.361; and after 8 h, 3.258 ± 0.094 vs 1.208 ± 0.082. For immunohistochemical staining in SAP vs SO, the results were: after 1 h, 1.793 ± 0.463 vs 0.808 ± 0.252; after 4 h, 4.535 ± 0.550 vs 0.871 ± 0.318; and after 8 h, 6.071 ± 0.941 vs 1.020 ± 0.406. Moreover, we observed a positive correlation in the pancreas (r = 0.852, P < 0.001) and PBMCs (r = 0.735, P < 0.001) between fgl2 expression and the severity of pancreatic injury. Masson staining showed that microthrombosis (%) in rats with SAP was increased (P < 0.001) compared with that in the SO group and it was closely correlated with fgl2 expression in the pancreas (r = 0.842, P < 0.001). For Masson staining in SAP vs SO, the results were: after 1 h, 26.880 ± 9.031 vs 8.630 ± 3.739; after 4 h, 53.750 ± 19.039 vs 8.500 ± 4.472; and after 8 h, 80.250 ± 12.915 vs 10.630 ± 7.003.CONCLUSION: Microthrombosis due to fgl2 overexpression contributes to pancreatic impairment in rats with SAP, and fgl2 level may serve as a biomarker during early stages of disease.

The development of blood-retinal barrier during the interaction of astrocytes with vascular wall cells

Astrocytes are intimately involved in the formation and development of retinal vessels. Astrocyte dysfunction is a major cause of blood-retinal barrier injury and other retinal vascular diseases. In this study, the development of the retinal vascular system and the formation of the blood-ret-inal barrier in mice were investigated using immunolfuorescence staining, gelatin-ink perfusion, and transmission electron microscopy. The results showed that the retinal vascular system of mice develops from the optic disc after birth, and radiates out gradually to cover the entire retina, taking the papilla optica as the center. First, the superifcial vasculature is formed on the inner retinal layer;then, the vasculature extends into the inner and outer edges of the retinal inner nuclear layer, forming the deep vasculature that is parallel to the superifcial vasculature. The blood-retinal barrier is mainly composed of endothelium, basal lamina and the end-feet of astrocytes, which become mature during mouse development. Initially, the naive endothelial cells were immature with few organelles and many microvilli. The basal lamina was uniform in thickness, and the glial end-feet surrounded the outer basal lamina incompletely. In the end, the blood-retinal barrier matures with smooth endothelia connected through tight junctions, rela-tively thin and even basal lamina, and relatively thin glial cell end-feet. These ifndings indicate that the development of the vasculature in the retina follows the rules of“center to periphery”and“superifcial layer to deep layers”. Its development and maturation are spatially and tempo-rally consistent with the functional performance of retinal neurons and photosensitivity. The blood-retinal barrier gradually becomes mature via the process of interactions between astro-cytes and blood vessel cells.

Effect of the Principle of Activating Blood Circulation to Break Stasis on GMP-140 and D_2 Dimer in Patients with Acute Cerebral Infarction

Objective: To explore the clinical efficacy of the principle of activating blood circulation to break stasis (ABCBS) and its influence on platelet membranous protein particle (GMP-140) and D 2 dimer (D-dimer) before and after treatment. Methods: Eighty-eight patients with blood stasis syndrome (BSS) of acute cerebral infarction (ACI) were randomly divided into two groups, both of which were treated with conventional treatment, i.e. with western medicine (WM), with Salvia injection added through intravenously dripping.One of the two groups was used as the control and the other group as the treated group who had ABCBS herbs orally taken in addition. The duration of treatment course for both groups was 3 weeks. Results: There were changes in both groups over clinical symptoms, nerve function deficit scoring and GMP-140, D-dimer, but the treated group showed significantly better than that of the control group, ( P <0.05). Conclusion: ABCBS principle could serve as an important auxiliary treating method for BSS of ACI, as it can effectively alter the blood of ACI patients which was viscous, condense, coagulant and aggregating.

Association of high expression in rat gastric mucosal heat shock protein 70 induced by moxibustion pretreatment with protection against stress injury

AIM:To study the effect of moxibustion on Zusanli or Liangmeng point on gastric mucosa injury in stress-induced ulcer rats and its correlation with the expression of heat shock protein 70 (HSP70). METHODS:Sixty healthy SD rats (30 males,30 females) were divided into control group,injury model group,Zushanli point group,Liangmeng point group. Stress gastric ulcer model was induced by binding cold stress method. Gastric mucosa ulcer injury (UI) index was calculated by Guth method. Gastric mucosa blood flow (GMBF) was recorded with a biological signal analyzer. Protein content and gene expression in gastric mucosal HSP70 were detected by immunohistochemistry (IHC) and reverse transcription polymerase chain reaction (RT-PCR). Thiobarbital method was used to determine malondialdehyde (MDA) content. Gastric mucosal endothelin (ET) and prostaglandin E2 (PGE2) were analyzed by radioimmunoassay. RESULTS:High gastric mucosal UI index,high HSP70 expression,low GMBF and PGF2,elevated MDA and ET were observed in gastric mucosa of rats subjected to cold stress. Moxibustion on Zusanli or Liangmeng point decreased rat gastric mucosal UI index,MDA and ET. Conversely,the expression of HSP70,GMBF,and PGE2 was elevated in gastric mucosa after pretreatment with moxibustion on Zusanli or Liangmeng point. The observed parameters were significantly different between Zusanli and Liangmeng points. CONCLUSION:Pretreatment with moxibustion on Zusanli or Liangmeng point protects gastric mucosa against stress injury. This protection is associated with the higher expression of HSP70 mRNA and protein,leading to release of PGE2 and inhibition of MDA and ET,impairment of gastric mucosal index.

Emerging blood exosome-based biomarkers for preclinical and clinical Alzheimer's disease:a meta-analysis and systematic review

Blood exosomes,which are extracellular vesicles secreted by living cells into the circulating blood,are regarded as a relatively noninvasive novel tool for monitoring brain physiology and disease states.An increasing number of blood cargo-loaded exosomes are emerging as potential biomarkers for preclinical and clinical Alzheimer's disease.Therefo re,we conducted a meta-analysis and systematic review of molecular biomarkers derived from blood exosomes to comprehensively analyze their diagnostic performance in preclinical Alzheimer's disease,mild cognitive impairment,and Alzheimer's disease.We performed a literature search in PubMed,Web of Science,Embase,and Cochrane Library from their inception to August 15,2020.The research subjects mainly included Alzheimer's disease,mild cognitive impairment,and preclinical Alzheimer's disease.We identified 34 observational studies,of which 15 were included in the quantitative analysis(Newcastle-Ottawa Scale score 5.87 points)and 19 were used in the qualitative analysis.The meta-analysis results showed that core biomarkers including Aβ_(1-42),P-T181-tau,P-S396-tau,and T-tau were increased in blood neuro nderived exosomes of preclinical Alzheimer's disease,mild cognitive impairment,and Alzheimer's disease patients.M olecules related to additional risk facto rs that are involved in neuroinflammation(C1q),metabolism disorder(P-S312-IRS-1),neurotrophic deficiency(HGF),vascular injury(VEGF-D),and autophagy-lysosomal system dysfunction(cathepsin D)were also increased.At the gene level,the differential expression of transc ription-related factors(REST)and microRNAs(miR-132)also affects RNA splicing,transport,and translation.These pathological changes contribute to neural loss and synaptic dysfunction.The data confirm that the above-mentioned core molecules and additional ris k-related factors in blood exosomes can serve as candidate biomarkers for preclinical and clinical Alzheimer's disease.These findings support further development of exosome biomarkers for a clinical blood test for Alzheimer's disease.This meta-analysis was registered at the International Prospective Register of Systematic Reviews(Registration No.CRD4200173498,28/04/2020).